TY - JOUR AU - Owen, D. J. AU - Davis, C. B. AU - Hartnell, R. D. AU - Madge, P. D. AU - Thomson, R. J. AU - Chong, A. K. AU - Coppel, R. L. AU - Itzstein, M. V. PY - 2007 TI - Synthesis and evaluation of galactofuranosyl N,N-dialkyl sulfenamides and sulfonamides as antimycobacterial agents N1 - Jan 27 JF - Bioorg Med Chem Lett SN - 0960-894X (Print) N1 - Synthesis and evaluation of galactofuranosyl N,N-dialkyl sulfenamides and sulfonamides as antimycobacterial agents N1 - 17303419 N1 - Journal article N1 - Eng N2 - The recent emergence of clinically oppressive superbugs, some with resistance to nearly all frontline drug therapies, has challenged our ability to combat such infectious organisms as Mycobacterium tuberculosis, the causative agent of tuberculosis (TB). Our medicinal chemistry program targeting this pathogen has identified several potent galactofuranose-based in vitro inhibitors of mycobacterial growth. The most potent compound, the Galf N,N-didecyl sulfenamide 8d, displayed anti-mycobacterial activity (MIC) of 1mug/mL in a cell based assay against a representative strain of Mycobacterium smegmatis. AD - Institute for Glycomics, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre, Qld. 9726, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17303419 ID - 3 ER - TY - JOUR AU - Oertelt, S. AU - Rieger, R. AU - Selmi, C. AU - Invernizzi, P. AU - Ansari, A. A. AU - Coppel, R. L. AU - Podda, M. AU - Leung, P. S. AU - Gershwin, M. E. PY - 2007 TI - A sensitive bead assay for antimitochondrial antibodies: Chipping away at AMA-negative primary biliary cirrhosis SP - 659-65 N1 - Mar JF - Hepatology JO - Hepatology (Baltimore, Md VL - 45 IS - 3 SN - 0270-9139 (Print) N1 - A sensitive bead assay for antimitochondrial antibodies: Chipping away at AMA-negative primary biliary cirrhosis N1 - 17326160 N1 - Dk39588/dk/niddk Dk92310/dk/niddk Journal Article Research Support, N.I.H., Extramural United States N1 - eng N2 - The antimitochondrial response in primary biliary cirrhosis (PBC) is the most highly directed and specific self-reacting antibody in human immunopathology. Originally, antimitochondrial antibodies (AMAs) were detected by indirect immunofluorescence (IIF) and found in approximately 90% of well-documented patients with PBC. The introduction of recombinant autoantigens and the use of immunoblotting have increased the sensitivity and specificity of AMAs, and they are now considered positive in approximately 95% of patients with PBC. Clearly, accurate autoantibody detection represents one of the fundamental requirements for reliable diagnostics in autoimmunity. To address the 5% of AMA-negative patients with PBC, we have generated and validated a bead assay for the detection of AMA. We enrolled 120 patients with PBC, including a non-random group of 30 rigorously proven AMA-negative patients, 50 healthy subjects, and 74 controls with autoimmune diseases (18 with primary sclerosing cholangitis, 16 with autoimmune hepatitis, and 40 with systemic lupus erythematosus). Individual bead assays were done with the three mitochondrial autoantigens, PDC-E2, BCOADC-E2, and OGDC-E2. As expected, 90 of 90 previously known AMA-positive patients remained positive with this assay but, interestingly, 20% of the rigorously defined AMA-negative patient group had antibodies to one or more of the mitochondrial autoantigens. Furthermore, 100% of these newly detected AMA-positive patients were anti-nuclear antibody (ANA) positive. CONCLUSION: The development of this assay reflects the potential for automated detection with rapid and reliable assaying and further highlights the diminished number of truly AMA-negative PBC patients. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17326160 ID - 1 ER - TY - JOUR AU - Moritoki, Y. AU - Lian, Z. X. AU - Wulff, H. AU - Yang, G. X. AU - Chuang, Y. H. AU - Lan, R. Y. AU - Ueno, Y. AU - Ansari, A. A. AU - Coppel, R. L. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 2007 TI - AMA production in primary biliary cirrhosis is promoted by the TLR9 ligand CpG and suppressed by potassium channel blockers SP - 314-22 N1 - Feb JF - Hepatology JO - Hepatology (Baltimore, Md VL - 45 IS - 2 SN - 0270-9139 (Print) N1 - AMA production in primary biliary cirrhosis is promoted by the TLR9 ligand CpG and suppressed by potassium channel blockers N1 - 17256753 N1 - Dk39588/dk/niddk Dk92310/dk/niddk Journal Article Research Support, N.I.H., Extramural United States N1 - eng N2 - We previously reported that peripheral blood mononuclear cells (PBMCs) from patients with primary biliary cirrhosis (PBC) produce significantly higher levels of polyclonal IgM than controls after exposure to CpG. Furthermore, the prevalence and unusually high levels of antimitochondrial antibodies (AMAs) in patients with PBC suggest a profound loss ofB cell tolerance. We have addressed the issue of whether CpG will promote the production ofAMAs and whether new experimental agents that inhibit the lymphocyte potassium channels Kv1.3 and KCa3.1 can suppress CpG-mediated B cell activation and AMA production. PBMCs were stimulated with and without CpG and were subsequently analyzed for phenotype, including expression of TLR9, CD86, and KCa3.1 concurrent with measurements of AMA and responses to a control antigen, tetanus toxoid, in supernatants. Additionally, K+ channel expression on B cells from PBC patients and controls was studied using whole-cell patch-clamp technology. In patients with PBC, CpG induces secretion of AMAs in PBMCs andalso up-regulates B cell expression of TLR9, CD86, and KCa3.1. Additionally, K+ channel blockers suppress secretion of AMA without a reduction of CpG-B-enhanced IgM production. Furthermore, there is diminished up-regulation of TLR9 and CD86 without affecting proliferation of B cells, B cell apoptosis, or viability. Conclusion: These data suggest that the hyperresponsiveness of B cells in PBC accelerates B cell-mediated autoimmunity. AD - Division of Rheumatology, Allergy, and Clinical Immunology and the University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17256753 ID - 4 ER - TY - JOUR AU - Logan, G. J. AU - Wang, L. AU - Zheng, M. AU - Cunningham, S. C. AU - Coppel, R. L. AU - Alexander, I. E. PY - 2007 TI - AAV vectors encoding malarial antigens stimulate antigen-specific immunity but do not protect from parasite infection SP - 1014-22 N1 - Jan 22 JF - Vaccine JO - Vaccine VL - 25 IS - 6 SN - 0264-410X (Print) N1 - AAV vectors encoding malarial antigens stimulate antigen-specific immunity but do not protect from parasite infection N1 - 17081661 N1 - Journal Article Netherlands N1 - eng N2 - This study explores the utility of recombinant adeno-associated virus (rAAV) as a genetic vaccine delivery system using muscle as a target tissue. A single injection of rAAV encoding the malarial antigens MSP4 (Plasmodium falciparum) or MSP4/5 (Plasmodium yoelii) stimulated long-term antigen-specific antibody responses. Anti-MSP4/5 immunity stimulated by AAV was not protective against P. yoelii infection and efforts taken to augment antibody responses against MSP4/5, either by priming with plasmid DNA or AAV and boosting with rAAV were unsuccessful. Alternative strategies such as inclusion of genetic adjuvants into the AAV vector will be necessary to stimulate an adequate level of anti-malarial protective immunity in this model. AD - Gene Therapy Research Unit, Children's Medical Research Institute and The Children's Hospital at Westmead, Westmead, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17081661 ID - 5 ER - TY - JOUR AU - Lea-Smith, D. J. AU - Pyke, J. S. AU - Tull, D. AU - McConville, M. J. AU - Coppel, R. L. AU - Crellin, P. K. PY - 2007 TI - The reductase that catalyzes mycolic motif synthesis is required for efficient attachment of mycolic acids to arabinogalactan N1 - Feb 17 JF - J Biol Chem SN - 0021-9258 (Print) N1 - The reductase that catalyzes mycolic motif synthesis is required for efficient attachment of mycolic acids to arabinogalactan N1 - 17308303 N1 - Journal article N1 - Eng N2 - Mycolic acids are essential components of the cell walls of bacteria belonging to the suborder Corynebacterineae, including the important human pathogens Mycobacterium tuberculosis and Mycobacterium leprae. Mycolic acid biosynthesis is complex and the target of several front-line antimycobacterial drugs. The condensation of two fatty acids to form a 2-alkyl-3-keto mycolate precursor, and the subsequent reduction of this precursor, represent two key and highly conserved steps in this pathway. While the enzyme catalyzing the condensation step has recently been identified, little is known about the putative reductase. Using an extensive bioinformatic comparison of the genomes of M. tuberculosis and Corynebacterium glutamicum we identified NCgl2385, the orthologue of Rv2509 in M. tuberculosis, as a potential reductase candidate. Deletion of the gene in C. glutamicum resulted in a slow growing strain that was deficient in arabinogalactan-linked mycolates and synthesized abnormal forms of the mycolate-containing glycolipids trehalose dicorynomycolate and trehalose monocorynomycolate. Analysis of the native and acetylated trehalose glycolipids by MALDI-TOF mass spectrometry indicated that these novel glycolipids contained an unreduced ss-keto-ester. This was confirmed by analysis of sodium borodeuteride reduced mycolic acids by gas chromatography mass spectrometry. Reintroduction of the NCgl2385 gene into the mutant restored the transfer of mature mycolic acids to both the trehalose glycolipids and cell wall arabinogalactan. These data indicate that NCgl2385, which we have designated CmrA, is essential for the production of mature trehalose mycolates and subsequent covalent attachment of mycolic acids onto the cell wall, thus representing a focus for future structural and pathogenicity studies. AD - Microbiology, Monash University, Monash University, Victoria 3800. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17308303 ID - 2 ER - TY - JOUR AU - Wakabayashi, K. AU - Lian, Z. X. AU - Moritoki, Y. AU - Lan, R. Y. AU - Tsuneyama, K. AU - Chuang, Y. H. AU - Yang, G. X. AU - Ridgway, W. AU - Ueno, Y. AU - Ansari, A. A. AU - Coppel, R. L. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 2006 TI - IL-2 receptor alpha(-/-) mice and the development of primary biliary cirrhosis SP - 1240-9 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 44 IS - 5 SN - 0270-9139 (Print) N1 - IL-2 receptor alpha(-/-) mice and the development of primary biliary cirrhosis N1 - 17058261 N1 - Dk39588/dk/niddk Journal Article Research Support, N.I.H., Extramural United States N1 - eng KW - Animals Cytokines/*blood Disease Models, Animal Female Flow Cytometry Immunoglobulins/*blood Interleukin-2 Receptor alpha Subunit/*deficiency/immunology Liver Cirrhosis, Biliary/*etiology/immunology Lymphocyte Count Male Mice Mice, Inbred C57BL Mitochondria/*immunology Phenotype T-Lymphocytes, Regulatory/*pathology N2 - Recently, we identified a child born with a genetic deficiency of IL-2 receptor alpha (IL-2Ralpha, CD25) expression who had several clinical manifestations of primary biliary cirrhosis (PBC). In addition, there has been suggestive evidence in both patients with PBC and their first-degree relatives that a deficiency of regulatory T cells (Tregs) is an integral component for susceptibility to PBC. Based on these observations, we generated IL-2Ralpha/CD25 deficient (IL-2Ralpha(-/-)) mice and wild-type littermate controls and followed them longitudinally for the natural history of liver immunopathology and appearance of antimitochondrial antibodies (AMAs). The analyses included immunohistochemical staining of liver and portal tract infiltrates as well as FACS profiles of lymphoid subpopulations in liver and spleen. In addition, serum cytokine profiles were quantitated. Importantly, IL-2Ralpha(-/-), but not littermate controls, develop portal inflammation and biliary ductular damage similar to human patients with PBC. CD4(+) and CD8(+) T cells predominate among portal cell infiltrates and sera reflect a Th1 cytokine bias with increased levels of IFN-gamma, TNF-alpha, IL-2 and IL-12p40. Of importance is the finding that the IL-2Ralpha(-/-) mice not only develop significantly increased serum levels of IgG and IgA, but they also develop AMAs with specificity for PDC-E2, which maps to the inner lipoyl domain of the autoantigen, all characteristics which are hallmarks of human PBC. In conclusion, the IL-2Ralpha(-/-) mice should facilitate studies of the early events in PBC and especially the tantalizing connection between Treg deficiency and autoimmunity specifically directed to mitochondrially located PDC-E2 and subsequent biliary ductular cell damage. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, Davis, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17058261 ID - 6 ER - TY - JOUR AU - Sanders, P. R. AU - Kats, L. M. AU - Drew, D. R. AU - O'Donnell, R. A. AU - O'Neill, M. AU - Maier, A. G. AU - Coppel, R. L. AU - Crabb, B. S. PY - 2006 TI - A set of glycosylphosphatidyl inositol-anchored membrane proteins of Plasmodium falciparum is refractory to genetic deletion SP - 4330-8 N1 - Jul JF - Infect Immun JO - Infection and immunity VL - 74 IS - 7 SN - 0019-9567 (Print) N1 - A set of glycosylphosphatidyl inositol-anchored membrane proteins of Plasmodium falciparum is refractory to genetic deletion N1 - 16790807 N1 - Dk 32094/dk/niddk Wellcome Trust Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States N1 - eng KW - Animals Antigens, Protozoan/chemistry/genetics/metabolism Cell Line *Gene Deletion Glycosylphosphatidylinositols/*chemistry/metabolism Membrane Proteins/chemistry/*genetics/metabolism Plasmodium falciparum/*genetics/metabolism/pathogenicity Protozoan Proteins/chemistry/*genetics/metabolism N2 - Targeted gene disruption has proved to be a powerful approach for studying the function of important ligands involved in erythrocyte invasion by the extracellular merozoite form of the human malaria parasite, Plasmodium falciparum. Merozoite invasion proceeds via a number of seemingly independent alternate pathways, such that entry can proceed with parasites lacking particular ligand-receptor interactions. To date, most focus in this regard has been on single-pass (type 1) membrane proteins that reside in the secretory organelles. Another class of merozoite proteins likely to include ligands for erythrocyte receptors are the glycosylphosphatidyl inositol (GPI)-anchored membrane proteins that coat the parasite surface and/or reside in the apical organelles. Several of these are prominent vaccine candidates, although their functions remain unknown. Here, we systematically attempted to disrupt the genes encoding seven of the known GPI-anchored merozoite proteins of P. falciparum by using a double-crossover gene-targeting approach. Surprisingly, and in apparent contrast to other merozoite antigen classes, most of the genes (six of seven) encoding GPI-anchored merozoite proteins are refractory to genetic deletion, with the exception being the gene encoding merozoite surface protein 5 (MSP-5). No distinguishable growth rate or invasion pathway phenotype was detected for the msp-5 knockout line, although its presence as a surface-localized protein was confirmed. AD - Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3050, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16790807 ID - 11 ER - TY - JOUR AU - Rieger, R. AU - Leung, P. S. AU - Jeddeloh, M. R. AU - Kurth, M. J. AU - Nantz, M. H. AU - Lam, K. S. AU - Barsky, D. AU - Ansari, A. A. AU - Coppel, R. L. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 2006 TI - Identification of 2-nonynoic acid, a cosmetic component, as a potential trigger of primary biliary cirrhosis SP - 7-16 N1 - Aug JF - J Autoimmun JO - Journal of autoimmunity VL - 27 IS - 1 SN - 0896-8411 (Print) N1 - Identification of 2-nonynoic acid, a cosmetic component, as a potential trigger of primary biliary cirrhosis N1 - 16876981 N1 - Dk037003/dk/niddk Dk39588/dk/niddk Journal Article Research Support, N.I.H., Extramural England N1 - eng KW - Autoantibodies/biosynthesis/*drug effects Cosmetics/adverse effects/*chemistry Fatty Acids, Unsaturated/*adverse effects/chemistry/*immunology Female Humans Liver Cirrhosis, Biliary/*chemically induced/immunology Male Mitochondrial Proteins/immunology Molecular Mimicry/immunology Occupational Exposure Structure-Activity Relationship N2 - Antimitochondrial antibodies (AMA) are unique among autoimmune serologic reactants because of their extremely high association with the index disease primary biliary cirrhosis (PBC). This autoantibody response is specifically directed only to the lipoyl domain of the mitochondrial 2-oxo-acid dehydrogenase complexes, which prompted us to search for environmental mimotopes in the form of xenobiotics and led to our identification of 2-octynoic acid as a high-affinity reactant for AMA. To focus on the chemical characteristics requisite for binding of AMA to the xenobiotic-modified self-peptide, quantitative structure-activity relationship (QSAR) studies were performed using a panel of alkynoic compounds, including examination of the length of the carbon chain and the location of the triple bond in the identified mimotope. Analyses of octynamides that varied in the position of the triple bond demonstrated that only the 2-octynamide reacted strongly with PBC sera. Furthermore, among 2-alkynamides with varying carbon chain length, 2-octyn-, 2-nonyn- (particularly) and 2-decynamide exhibited the highest reactivity. Thus, an optimal chemical structure of the xenobiotically modified epitope recognized by AMA-positive PBC sera is provided by 2-nonynoic acid. The methyl ester of this compound is ranked 2,324th out of 12,945 compounds to which there is occupational exposure, with an 80% female prevalence due to its use in cosmetic products. Our findings illustrate an unusual polyreactivity of anti-PDC-E2 and support the idea of epitope mimicry in the genesis of this autoantibody and perhaps of PBC itself. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, GBSF 6510, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16876981 ID - 9 ER - TY - JOUR AU - Plebanski, M. AU - Lopez, E. AU - Proudfoot, O. AU - Cooke, B. M. AU - Itzstein, M. AU - Coppel, R. L. PY - 2006 TI - Economic and practical challenges to the formulation of vaccines against endemic infectious diseases such as malaria SP - 77-85 N1 - Sep JF - Methods JO - Methods (San Diego, Calif VL - 40 IS - 1 SN - 1046-2023 (Print) N1 - Economic and practical challenges to the formulation of vaccines against endemic infectious diseases such as malaria N1 - 16997716 N1 - Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States N1 - eng KW - Chemistry, Pharmaceutical/*economics/*methods/trends Communicable Disease Control/*economics Malaria Vaccines/*economics Vaccines/chemistry/*economics N2 - Herein, we analyze in general the current vaccine market and identify potential factors driving and modulating supply and demand for vaccines. An emphasis is placed on changes in regulation in the last 20 years which have led to increased indirect costs of production, and which can create a barrier against the timely use of technological advances to reduce direct costs. Other defining industry characteristics, such as firm numbers and sizes, cost and pricing strategies, nature extent and impact of Government involvement and international regulation are noted. These considerations, far from being removed from basic vaccine research, influence its ability to achieve aims that can be then progressed into effective vaccine products. We discuss specifically the development of particulate vaccines against malaria, a major lethal disease and health problem prevalent in Africa, including some key economic and methodological challenges and opportunities. We note some practical issues blocking the development of effective particulate vaccines for the Third World, mainly driven by the regulatory spiral noted above. AD - Vaccine and Infectious Diseases Laboratory, Burnet Institute at Austin, Studley Road, Heidelberg, Vic. 3084, Australia. mplebans@burnet.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16997716 ID - 7 ER - TY - JOUR AU - Oertelt, S. AU - Lian, Z. X. AU - Cheng, C. M. AU - Chuang, Y. H. AU - Padgett, K. A. AU - He, X. S. AU - Ridgway, W. M. AU - Ansari, A. A. AU - Coppel, R. L. AU - Li, M. O. AU - Flavell, R. A. AU - Kronenberg, M. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 2006 TI - Anti-mitochondrial antibodies and primary biliary cirrhosis in TGF-beta receptor II dominant-negative mice SP - 1655-60 N1 - Aug 1 JF - J Immunol VL - 177 IS - 3 SN - 0022-1767 (Print) N1 - Anti-mitochondrial antibodies and primary biliary cirrhosis in TGF-beta receptor II dominant-negative mice N1 - 16849474 N1 - Dk39588/dk/niddk Journal Article Research Support, N.I.H., Extramural United States 1950) N1 - eng KW - Animals Autoantibodies/*biosynthesis Bile Ducts/immunology/pathology CD4-Positive T-Lymphocytes/pathology CD8-Positive T-Lymphocytes/pathology Cell Movement/genetics/immunology Cytokines/biosynthesis Female Liver/immunology/pathology Liver Cirrhosis, Biliary/*genetics/*immunology/pathology Lymphocyte Subsets/pathology Male Mice Mice, Inbred C57BL Mice, Knockout Mice, Transgenic Mitochondria/*immunology Receptors, Transforming Growth Factor beta/*deficiency/*genetics Species Specificity N2 - Primary biliary cirrhosis (PBC) is an autoimmune disease of the liver, characterized by lymphocytic infiltrates in portal tracts, selective destruction of biliary epithelial cells, and anti-mitochondrial Abs (AMAs). The elucidation of early events in the induction of tissue inflammation and autoimmunity in PBC has been hampered by the cryptic onset of the disease, the practical limitations in accessing the target tissue, and the lack of a suitable animal model. We demonstrate in this study that a mouse transgenic for directed expression of a dominant-negative form of TGF-beta receptor type II (dnTGFbetaRII), under the direction of the CD4 promoter, mimics several key phenotypic features of human PBC, including spontaneous production of AMAs directed to the same mitochondrial autoantigens, namely PDC-E2, BCOADC-E2, and OGDC-E2. The murine AMAs also inhibit PDC-E2 activity. Moreover, there is lymphocytic liver infiltration with periportal inflammation analogous to the histological profile in human PBC. Additionally, the serum cytokine profile of affected mice mimics data in human PBC. The concomitant presence of these immunopathological features in the transgenic mice suggests that the TGF-betaRII pathway is implicated in the pathogenesis of PBC. Finally, these data point away from initiation of autoimmunity by mechanisms such as molecular mimicry and more toward activation of an intrinsically self-reactive T cell repertoire in which necessary regulatory T cell influences are lacking. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16849474 ID - 10 ER - TY - JOUR AU - Marland, Z. AU - Beddoe, T. AU - Zaker-Tabrizi, L. AU - Lucet, I. S. AU - Brammananth, R. AU - Whisstock, J. C. AU - Wilce, M. C. AU - Coppel, R. L. AU - Crellin, P. K. AU - Rossjohn, J. PY - 2006 TI - Hijacking of a substrate-binding protein scaffold for use in mycobacterial cell wall biosynthesis SP - 983-97 N1 - Jun 16 JF - J Mol Biol JO - Journal of molecular biology VL - 359 IS - 4 SN - 0022-2836 (Print) N1 - Hijacking of a substrate-binding protein scaffold for use in mycobacterial cell wall biosynthesis N1 - 16698034 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Bacterial Proteins/*chemistry/*metabolism Binding Sites Cell Wall/*metabolism Crystallography, X-Ray Lipopolysaccharides/biosynthesis Models, Molecular Molecular Sequence Data Phosphatidylinositols/metabolism Phylogeny Protein Conformation Sequence Homology, Amino Acid Structural Homology, Protein N2 - The waxy cell wall is crucial to the survival of mycobacteria within the infected host. The cell wall is a complex structure rich in unusual molecules that includes two related lipoglycans, the phosphatidylinositol mannosides (PIMs) and lipoarabinomannans (LAMs). Many proteins implicated in the PIM/LAM biosynthetic pathway, while attractive therapeutic targets, are poorly defined. The 2.4A resolution crystal structure of an essential lipoprotein, LpqW, implicated in LAM biosynthesis is reported here. LpqW adopts a scaffold reminiscent of the distantly related, promiscuous substrate-binding proteins of the ATP-binding cassette import system. Nevertheless, the unique closed conformation of LpqW suggests that mycobacteria and other closely related pathogens have hijacked this scaffold for use in key processes of cell wall biosynthesis. In silico docking provided a plausible model in which the candidate PIM ligand binds within a marked electronegative region located on the surface of LpqW. We suggest that LpqW represents an archetypal lipoprotein that channels intermediates from a pathway for mature PIM production into a pathway for LAM biosynthesis, thus controlling the relative abundance of these two important components of the cell wall. AD - The Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Vic. 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16698034 ID - 14 ER - TY - JOUR AU - Lan, R. Y. AU - Cheng, C. AU - Lian, Z. X. AU - Tsuneyama, K. AU - Yang, G. X. AU - Moritoki, Y. AU - Chuang, Y. H. AU - Nakamura, T. AU - Saito, S. AU - Shimoda, S. AU - Tanaka, A. AU - Bowlus, C. L. AU - Takano, Y. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2006 TI - Liver-targeted and peripheral blood alterations of regulatory T cells in primary biliary cirrhosis SP - 729-37 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 43 IS - 4 SN - 0270-9139 (Print) N1 - Liver-targeted and peripheral blood alterations of regulatory T cells in primary biliary cirrhosis N1 - 16557534 N1 - Dk39588/dk/niddk Dk92310/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States N1 - eng KW - *Autoimmunity CD4 Lymphocyte Count CD4-Positive T-Lymphocytes/metabolism/pathology Case-Control Studies Forkhead Transcription Factors/metabolism Hepatitis C, Chronic/blood/immunology Hepatitis, Autoimmune/blood/immunology Humans Liver/*immunology Liver Cirrhosis, Biliary/*blood/*immunology/pathology/physiopathology Receptors, Interleukin-2/metabolism Receptors, Nerve Growth Factor/metabolism Receptors, Tumor Necrosis Factor/metabolism Severity of Illness Index T-Lymphocytes, Regulatory/*immunology/metabolism/pathology N2 - CD4+CD25high regulatory T cells (Tregs) play a critical role in self-tolerance, as seen in murine autoimmunity. Studies on Tregs in human autoimmunity have focused primarily on peripheral blood samples. A study targeting diseased tissue should identify direct relationships between Tregs and autoimmunity. Peripheral blood samples were collected from 91 patients with primary biliary cirrhosis (PBC), 28 immediate relatives, and 41 healthy controls, and Treg frequencies were determined as a percentage of CD4+CD25high T cells in CD4+TCR-alphabeta+ T cells. A tissue-targeted determination of frequency and distribution of FoxP3+ Tregs was also performed on 90 different liver tissue specimens exhibiting PBC (n = 52), chronic hepatitis C (CHC) (n = 30), and autoimmune hepatitis (AIH) (n = 8). Treg suppression studies were performed on 50 PBC patients and 27 controls. Patients with PBC demonstrated a relative reduction of Tregs compared with controls (P < .0002). Interestingly, a deficiency in CD4+CD25+ Tregs was also found in the daughters and sisters of PBC patients compared with controls (P < .0007). However, functional studies did not reveal a global PBC Treg defect. The level of FoxP3-expressing Tregs was markedly lower in affected PBC portal tracts compared with CHC and AIH (P < .001). In addition, the CD8+T cell/FoxP3+ Treg ratio was significantly higher in livers of late-stage PBC compared with those of CHC (P < .001) and early-stage AIH (P < .001). In conclusion, these data provide support for a genetic modulation of Treg frequency and illustrate the role Tregs play in the loss of tolerance in PBC. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California--Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16557534 ID - 16 ER - TY - JOUR AU - Kovacevic, S. AU - Anderson, D. AU - Morita, Y. S. AU - Patterson, J. AU - Haites, R. AU - McMillan, B. N. AU - Coppel, R. AU - McConville, M. J. AU - Billman-Jacobe, H. PY - 2006 TI - Identification of a novel protein with a role in lipoarabinomannan biosynthesis in mycobacteria SP - 9011-7 N1 - Apr 7 JF - J Biol Chem JO - The Journal of biological chemistry VL - 281 IS - 14 SN - 0021-9258 (Print) N1 - Identification of a novel protein with a role in lipoarabinomannan biosynthesis in mycobacteria N1 - 16455649 N1 - Journal Article Research Support, Non-U.S. Gov't United States N1 - eng KW - Cell Membrane/physiology Gene Expression Regulation, Bacterial *Genes, Bacterial Lipopolysaccharides/*biosynthesis Mutation Mycobacterium smegmatis/*genetics/growth & development/pathogenicity/*physiology Phenotype Phosphatidylinositols/*metabolism Virulence N2 - All species of Mycobacteria synthesize distinctive cell walls that are rich in phosphatidylinositol mannosides (PIMs), lipomannan (LM), and lipoarabinomannan (LAM). PIM glycolipids, having 2-4 mannose residues, can either be channeled into polar PIM species (with 6 Man residues) or hypermannosylated to form LM and LAM. In this study, we have identified a Mycobacterium smegmatis gene, termed lpqW, that is required for the conversion of PIMs to LAM and is highly conserved in all mycobacteria. A transposon mutant, Myco481, containing an insertion near the 3' end of lpqW exhibited altered colony morphology on complex agar medium. This mutant was unstable and was consistently overgrown by a second mutant, represented by Myco481.1, that had normal growth and colony characteristics. Biochemical analysis and metabolic labeling studies showed that Myco481 synthesized the complete spectrum of apolar and polar PIMs but was unable to make LAM. LAM biosynthesis was restored to near wild type levels in Myco481.1. However, this mutant was unable to synthesize the major polar PIM (AcPIM6) and accumulated a smaller intermediate, AcPIM4. Targeted disruption of the lpqW gene and complementation of the initial Myco481 mutant with the wild type gene confirmed that the phenotype of this mutant was due to loss of LpqW. These studies suggest that LpqW has a role in regulating the flux of early PIM intermediates into polar PIM or LAM biosynthesis. They also suggest that AcPIM4 is the likely branch point intermediate in polar PIM and LAM biosynthesis. AD - ARC Centre for Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16455649 ID - 19 ER - TY - JOUR AU - Kats, L. M. AU - Black, C. G. AU - Proellocks, N. I. AU - Coppel, R. L. PY - 2006 TI - Plasmodium rhoptries: how things went pear-shaped SP - 269-76 N1 - Jun JF - Trends Parasitol JO - Trends in parasitology VL - 22 IS - 6 SN - 1471-4922 (Print) N1 - Plasmodium rhoptries: how things went pear-shaped N1 - 16635585 N1 - Journal Article Research Support, Non-U.S. Gov't Review England N1 - eng KW - Animals Erythrocytes/*parasitology Host-Parasite Relations Humans Organelles/*physiology/ultrastructure Plasmodium/pathogenicity/*physiology/ultrastructure Protozoan Proteins/metabolism/*physiology/ultrastructure N2 - Plasmodium parasites have three sets of specialised secretory organelles at the apical end of their invasive forms--rhoptries, micronemes and dense granules. The contents of these organelles are responsible for or contribute to host cell invasion and modification, and at least four apical proteins are leading vaccine candidates. Given the unusual nature of Plasmodium invasion, it is not surprising that unique proteins are involved in this process. Nowhere is this more evident than in rhoptries. We have collated data from several recent studies to compile a rhoptry proteome. Discussion is focussed here on rhoptry content and function. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16635585 ID - 15 ER - TY - JOUR AU - He, X. S. AU - Ansari, A. A. AU - Ridgway, W. M. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2006 TI - New insights to the immunopathology and autoimmune responses in primary biliary cirrhosis SP - 1-13 N1 - Jan JF - Cell Immunol JO - Cellular immunology VL - 239 IS - 1 SN - 0008-8749 (Print) N1 - New insights to the immunopathology and autoimmune responses in primary biliary cirrhosis N1 - 16765923 N1 - Journal Article Review United States N1 - eng KW - Adaptation, Biological/immunology Animals Autoimmunity/*immunology Humans Immunity, Natural/immunology Liver Cirrhosis, Biliary/blood/etiology/*immunology/*pathology Molecular Mimicry/immunology T-Lymphocytes/immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease with profound changes in different compartments of the immune system, including those involved in innate, and adaptive immunity. New data from epidemiological studies of PBC have reinforced the thesis that the cause for this relatively uncommon disease is likely to be a combination of both environmental factors and a susceptible genetic predisposition. Recent findings of abnormalities of the innate immune system in PBC suggest that they may serve as links between the environmental factors and the early events in PBC development. Viral and bacterial infections as well as xenobiotics are some of the potential environmental factors that have been implicated in this complex process. Identification of the etiological factors for PBC will point to new preventive or therapeutic treatments. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California, Davis, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16765923 ID - 12 ER - TY - JOUR AU - Cooke, B. M. AU - Buckingham, D. W. AU - Glenister, F. K. AU - Fernandez, K. M. AU - Bannister, L. H. AU - Marti, M. AU - Mohandas, N. AU - Coppel, R. L. PY - 2006 TI - A Maurer's cleft-associated protein is essential for expression of the major malaria virulence antigen on the surface of infected red blood cells SP - 899-908 N1 - Mar 13 JF - J Cell Biol JO - The Journal of cell biology VL - 172 IS - 6 SN - 0021-9525 (Print) N1 - A Maurer's cleft-associated protein is essential for expression of the major malaria virulence antigen on the surface of infected red blood cells N1 - 16520384 N1 - 069515/Wellcome Trust Ai44008/ai/niaid Dk32094/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States N1 - eng KW - Animals Antigens, Surface/*metabolism Carrier Proteins/*genetics Cell Adhesion/genetics Down-Regulation/genetics Erythrocyte Membrane/genetics/metabolism/ultrastructure Erythrocytes/metabolism/*parasitology/ultrastructure Exocytosis/genetics Gene Expression Regulation/physiology Humans Membrane Proteins/*genetics Microscopy, Electron, Scanning Microscopy, Electron, Transmission Organelles/*metabolism/ultrastructure Plasmodium falciparum/genetics/*metabolism/ultrastructure Protein Transport/genetics Protozoan Proteins/*genetics/*metabolism N2 - The high mortality of Plasmodium falciparum malaria is the result of a parasite ligand, PfEMP1 (P. falciparum) erythrocyte membrane protein 1), on the surface of infected red blood cells (IRBCs), which adheres to the vascular endothelium and causes the sequestration of IRBCs in the microvasculature. PfEMP1 transport to the IRBC surface involves Maurer's clefts, which are parasite-derived membranous structures in the IRBC cytoplasm. Targeted gene disruption of a Maurer's cleft protein, SBP1 (skeleton-binding protein 1), prevented IRBC adhesion because of the loss of PfEMP1 expression on the IRBC surface. PfEMP1 was still present in Maurer's clefts, and the transport and localization of several other Maurer's cleft proteins were unchanged. Maurer's clefts were altered in appearance and were no longer found as close to the periphery of the IRBC. Complementation of mutant parasites with sbp1 led to the reappearance of PfEMP1 on the IRBC surface and the restoration of adhesion. Our results demonstrate that SBP1 is essential for the translocation of PfEMP1 onto the surface of IRBCs and is likely to play a pivotal role in the pathogenesis of P. falciparum malaria. AD - Molecular and Cellular Rheology Laboratory, Department of Microbiology, Monash University, Victoria 3800, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16520384 ID - 17 ER - TY - JOUR AU - Chuang, Y. H. AU - Lian, Z. X. AU - Tsuneyama, K. AU - Chiang, B. L. AU - Ansari, A. A. AU - Coppel, R. L. AU - Eric Gershwin, M. PY - 2006 TI - Increased killing activity and decreased cytokine production in NK cells in patients with primary biliary cirrhosis SP - 232-40 N1 - Jun JF - J Autoimmun JO - Journal of autoimmunity VL - 26 IS - 4 SN - 0896-8411 (Print) N1 - Increased killing activity and decreased cytokine production in NK cells in patients with primary biliary cirrhosis N1 - 16730427 N1 - Dk39588/dk/niddk N01-dk-9-2310/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't England N1 - eng KW - Adult Aged Cytokines/*biosynthesis/blood/immunology Female Humans Immunity, Natural/immunology Interferon Type II/blood/immunology Interleukin-6/blood/immunology Interleukin-8/biosynthesis/blood/immunology Killer Cells, Natural/*immunology Liver Cirrhosis, Biliary/blood/*immunology Male Membrane Glycoproteins/biosynthesis/blood/immunology Middle Aged Poly I-C/pharmacology Pore Forming Cytotoxic Proteins Receptors, Interleukin-8A/biosynthesis/blood/immunology N2 - Although the pathogenesis of primary biliary cirrhosis (PBC) remains enigmatic, the immune system plays a key role in the initiation and subsequent development of pathology. Previous studies have indicated a critical role of the innate immune system. Importantly, natural killer (NK) cells are abundant in liver where they serve as sentinels of the immune system. In addition, NK cells have significant biologic activity based on their production of immunoregulatory cytokines. To address this issue, we have investigated several qualitative and quantitative activities of NK cells in patients with PBC as well as normal and liver diseased controls. We report herein a marked increase in the frequency and absolute number of blood and liver NK cells in PBC patients. Moreover, the cytotoxic activity and perforin expression by isolated NK cells were significantly increased in PBC patients associated with increased levels of plasma IL-8 and the expression of CD128a (IL-8 receptor) on NK cells. In contrast, the levels of IFN-gamma, IL-6 and IL-8 synthesized by NK cells were significantly decreased in PBC patients as compared to controls. In conclusion, data from this study provide compelling evidence supporting a biologic role of NK cells in the immunopathogenesis of PBC. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis School of Medicine, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16730427 ID - 13 ER - TY - JOUR AU - Bulach, D. M. AU - Zuerner, R. L. AU - Wilson, P. AU - Seemann, T. AU - McGrath, A. AU - Cullen, P. A. AU - Davis, J. AU - Johnson, M. AU - Kuczek, E. AU - Alt, D. P. AU - Peterson-Burch, B. AU - Coppel, R. L. AU - Rood, J. I. AU - Davies, J. K. AU - Adler, B. PY - 2006 TI - Genome reduction in Leptospira borgpetersenii reflects limited transmission potential SP - 14560-5 N1 - Sep 26 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 103 IS - 39 SN - 0027-8424 (Print) N1 - Genome reduction in Leptospira borgpetersenii reflects limited transmission potential N1 - 16973745 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United States N1 - eng KW - Animals Bacterial Proteins/secretion Cattle Chromosomes, Bacterial/genetics DNA Transposable Elements/genetics *Disease Transmission Gene Expression Regulation, Bacterial Genes, Bacterial/genetics Genome, Bacterial/*genetics Genomics Humans Leptospira/classification/*genetics/pathogenicity Leptospira interrogans/genetics Membrane Proteins/metabolism Molecular Sequence Data Replicon/genetics Virulence N2 - Leptospirosis is one of the most common zoonotic diseases in the world, resulting in high morbidity and mortality in humans and affecting global livestock production. Most infections are caused by either Leptospira borgpetersenii or Leptospira interrogans, bacteria that vary in their distribution in nature and rely on different modes of transmission. We report the complete genomic sequences of two strains of L. borgpetersenii serovar Hardjo that have distinct phenotypes and virulence. These two strains have nearly identical genetic content, with subtle frameshift and point mutations being a common form of genetic variation. Starkly limited regions of synteny are shared between the large chromosomes of L. borgpetersenii and L. interrogans, probably the result of frequent recombination events between insertion sequences. The L. borgpetersenii genome is approximately 700 kb smaller and has a lower coding density than L. interrogans, indicating it is decaying through a process of insertion sequence-mediated genome reduction. Loss of gene function is not random but is centered on impairment of environmental sensing and metabolite transport and utilization. These features distinguish L. borgpetersenii from L. interrogans, a species with minimal genetic decay and that survives extended passage in aquatic environments encountering a mammalian host. We conclude that L. borgpetersenii is evolving toward dependence on a strict host-to-host transmission cycle. AD - Australian Bacterial Pathogenesis Program, Victorian Bioinformatics Consortium, Department of Microbiology, Monash University, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16973745 ID - 8 ER - TY - JOUR AU - Wang, L. AU - Kedzierski, L. AU - Schofield, L. AU - Coppel, R. L. PY - 2005 TI - Influence of glycosylphosphatidylinositol anchorage on the efficacy of DNA vaccines encoding Plasmodium yoelii merozoite surface protein 4/5 SP - 4120-7 N1 - Jul 14 JF - Vaccine JO - Vaccine VL - 23 IS - 32 SN - 0264-410X (Print) N1 - Influence of glycosylphosphatidylinositol anchorage on the efficacy of DNA vaccines encoding Plasmodium yoelii merozoite surface protein 4/5 N1 - 15964480 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antibodies, Protozoan/biosynthesis/immunology Antigens, Protozoan/biosynthesis/genetics/*immunology/isolation & purification COS Cells Glycosylphosphatidylinositols/*metabolism Membrane Proteins/genetics/immunology Plasmodium yoelii/genetics/*immunology Protozoan Proteins/*genetics/immunology Vaccines, DNA/genetics/*immunology Vaccines, Subunit/genetics/immunology N2 - Immune responses induced to DNA vaccination vary considerably and depend on a variety of factors, including the physical form in which the antigen is expressed by target cells and presented to the immune system. Data on the effect of these factors will aid improved design of DNA vaccines and facilitate their further development. We examined the effect of different forms of surface anchoring on the immunogenicity of a DNA vaccine. A number of constructs were generated encoding Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5) with or without its C-terminal glycosylphosphatidylinositol (GPI) attachment signal, replacing the endogenous GPI signal of PyMSP4/5 with that of mouse decay-accelerating factor (DAF), a well-established model for GPI-anchoring in mammalian cells, or the transmembrane anchor and cytoplasmic tail of mouse tissue factor (TF). All constructs were demonstrated to express the full-length PyMSP4/5 in transfected COS cells and induce PyMSP4/5-specific antibodies in mice. The GPI attachment signal of PyMSP4/5 was found to function poorly in mammalian cells and result in a much lower level of PyMSP4/5 expression in vitro than its mammalian counterpart. The DNA vaccine containing the mammalian GPI attachment signal induced the highest levels of antibodies and impacted Ig isotype distribution, consistent with the presence of a CD1-restricted pathway of Ig formation to GPI-anchored membrane proteins. Despite the induction of specific antibodies, none of these DNA vaccines induced sufficient levels of antibodies to protect mice against a lethal challenge with P. yoelii. AD - Department of Microbiology and The Victoria Bioinformatics Consortium, Monash University, Clayton, Vic. 3800, Australia. lina.wang@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15964480 ID - 29 ER - TY - JOUR AU - Pouniotis, D. S. AU - Proudfoot, O. AU - Bogdanoska, V. AU - Scalzo, K. AU - Kovacevic, S. AU - Coppel, R. L. AU - Plebanski, M. PY - 2005 TI - Selectively impaired CD8+ but not CD4+ T cell cycle arrest during priming as a consequence of dendritic cell interaction with plasmodium-infected red cells SP - 3525-33 N1 - Sep 15 JF - J Immunol VL - 175 IS - 6 SN - 0022-1767 (Print) N1 - Selectively impaired CD8+ but not CD4+ T cell cycle arrest during priming as a consequence of dendritic cell interaction with plasmodium-infected red cells N1 - 16148095 N1 - Journal Article Research Support, Non-U.S. Gov't United States 1950) N1 - eng KW - Animals *Antigen Presentation CD4-Positive T-Lymphocytes/immunology/pathology CD8-Positive T-Lymphocytes/immunology/pathology Cell Communication/immunology *Cell Cycle Dendritic Cells/immunology/parasitology/*pathology Erythrocytes/immunology/*parasitology Interleukin-10/pharmacology Interleukins/pharmacology Malaria/blood/immunology Mice Plasmodium/*immunology T-Lymphocytes/immunology/*pathology N2 - Individuals living in malaria-endemic areas show generally low T cell responses to malaria Ags. In this study, we show murine dendritic cell (DC) interaction with parasitized erythrocytes (pRBC) arrested their maturation, resulting in impaired ability to stimulate naive, but not recall T cell responses in vitro and in vivo. Moreover, within the naive T cell population, pRBC-treated DC were selectively deficient in priming CD8(+) but not CD4(+) T cells. Indeed, DC that had taken up pRBC were shown for the first time to efficiently prime CD4(+) T cell responses to a known protective merozoite Ag, MSP4/5. In contrast, impaired priming resulted in decreases in both proliferation and cytokine production by CD8(+) T cells. Deficient priming was observed to both a model and a Plasmodium berghei-specific CD8(+) T cell epitope. The mechanisms underlying the inability of parasite-treated DC to prime CD8(+) T cells were explored. pRBC treatment of DC from wild-type C57BL/6, but not from IL-10 knockout animals, suppressed DC-mediated T cell priming across a Transwell, suggesting active IL-10-dependent suppression. CD8(+) T cells were arrested at the G(0) stage of the cell cycle after two cell divisions post-Ag stimulation. The proliferation arrest was partially reversible by the addition of IL-2 or IL-7 to responder cultures. These results suggest that in malaria-endemic areas, priming of CD8(+) T cell responses may be more difficult to induce via vaccination than the priming of CD4(+) T cells. Moreover, pathogens may selectively target the CD8(+) T cell arm of protective immunity for immune evasion. AD - Vaccine and Infectious Diseases Unit, The Austin Research Institute, Heidelberg, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16148095 ID - 23 ER - TY - JOUR AU - Pei, X. AU - An, X. AU - Guo, X. AU - Tarnawski, M. AU - Coppel, R. AU - Mohandas, N. PY - 2005 TI - Structural and functional studies of interaction between Plasmodium falciparum knob-associated histidine-rich protein (KAHRP) and erythrocyte spectrin SP - 31166-71 N1 - Sep 2 JF - J Biol Chem JO - The Journal of biological chemistry VL - 280 IS - 35 SN - 0021-9258 (Print) N1 - Structural and functional studies of interaction between Plasmodium falciparum knob-associated histidine-rich protein (KAHRP) and erythrocyte spectrin N1 - 16006556 N1 - Dk 26263/dk/niddk Dk 32094/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence Animals Cell Membrane/chemistry/metabolism Erythrocytes/cytology/*metabolism/parasitology Humans *Membrane Proteins/genetics/metabolism Peptide Fragments/genetics/metabolism *Peptides/genetics/metabolism Plasmodium falciparum/*metabolism *Protozoan Proteins/genetics/metabolism Recombinant Fusion Proteins/genetics/metabolism *Spectrin/genetics/metabolism Surface Plasmon Resonance N2 - Plasmodium falciparum dramatically modifies the structure and function of the membrane of the parasitized host erythrocyte. Altered membrane properties are the consequence of the interaction of a group of exported malaria proteins with host cell membrane proteins. KAHRP (the knob-associated histidine-rich protein), a member of this group, has been shown to interact with erythrocyte membrane skeletal protein spectrin. However, the molecular basis for this interaction has yet to be defined. In the present study, we defined the binding motifs in both KAHRP and spectrin and identified a functional role for this interaction. We showed that spectrin bound to a 72-amino-acid KAHRP fragment (residues 370-441). Among nine-spectrin fragments, which encompass the entire alpha and beta spectrin molecules (four alpha spectrin and five beta spectrin fragments), KAHRP bound only to one, the alpha N-5 fragment. The KAHRP-binding site within the alpha N-5 fragment was localized uniquely to repeat 4. The interaction of full-length spectrin dimer to KAHRP was inhibited by repeat 4 of alpha spectrin. Importantly, resealing of this repeat peptide into erythrocytes mislocalized KAHRP in the parasitized cells. We concluded that the interaction of KAHRP with spectrin is critical for appropriate membrane localization of KAHRP in parasitized erythrocytes. As the presence of KAHRP at the erythrocyte membrane is necessary for cytoadherence in vivo, our findings have implications for the development of new therapies for mitigating the severity of malaria infection. AD - Red Cell Physiology Laboratory, New York Blood Center, New York, New York, 10021, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16006556 ID - 28 ER - TY - JOUR AU - Padgett, K. A. AU - Selmi, C. AU - Kenny, T. P. AU - Leung, P. S. AU - Balkwill, D. L. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2005 TI - Phylogenetic and immunological definition of four lipoylated proteins from Novosphingobium aromaticivorans, implications for primary biliary cirrhosis SP - 209-19 N1 - May JF - J Autoimmun JO - Journal of autoimmunity VL - 24 IS - 3 SN - 0896-8411 (Print) N1 - Phylogenetic and immunological definition of four lipoylated proteins from Novosphingobium aromaticivorans, implications for primary biliary cirrhosis N1 - 15848043 N1 - Journal Article England N1 - eng KW - Acyltransferases/genetics/immunology Amino Acid Sequence Animals Antigens, Bacterial/*genetics/immunology Autoantigens/genetics/immunology Autoimmune Diseases/genetics/immunology Bacterial Proteins/*genetics/immunology Dihydrolipoyllysine-Residue Acetyltransferase Evolution, Molecular Humans Lipoproteins/*genetics/immunology Liver Cirrhosis, Biliary/genetics/immunology Molecular Mimicry/genetics/immunology Molecular Sequence Data Phylogeny Pyruvate Dehydrogenase Complex/genetics/immunology *Sequence Homology, Amino Acid Sphingomonadaceae/*genetics/immunology Thioctic Acid/genetics/immunology N2 - Novosphingobium aromaticivorans, a unique ubiquitous bacterium that metabolizes xenobiotics and activates environmental estrogens, has been suggested as a pathogenic factor in the development of primary biliary cirrhosis (PBC). To define the molecular basis of PBC sera reactivity, we investigated the characteristic of the bacterial antigens involved. We cloned and sequenced four genes from N. aromaticivorans coding for immunoreactive proteins, arbitrarily named Novo 1 through Novo 4. We subsequently analyzed these proteins for their homology to known mitochondrial proteins and defined their reactivity using monoclonal antibodies (mAbs), rabbit anti-lipoic acid antibody, and PBC/control sera. Moreover, we studied their phylogenetic relation with the known PBC autoantigens. Novo proteins have an extraordinary degree of amino acid homology with all of the major human mitochondrial autoantigens PDC-E2 (Novo 1 and 2), OGDC-E2 (Novo 3), and BCOADC-E2 (Novo 4). Moreover, Novo 1-4 contain a lipoylated domain, are recognized by AMA-positive sera, and react with specific mAbs to mitochondrial antigens. Interestingly, the phylogenetic relation of the proteins emphasizes the conservation of the lipoylated domain. In conclusion, our data provide a high degree of confidence that N. aromaticivorans may potentiate the breakdown of self tolerance in genetically susceptible individuals. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, GBSF, 451 E. Health Sciences Drive, Suite 6510, Davis, California 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15848043 ID - 31 ER - TY - JOUR AU - Matsoso, L. G. AU - Kana, B. D. AU - Crellin, P. K. AU - Lea-Smith, D. J. AU - Pelosi, A. AU - Powell, D. AU - Dawes, S. S. AU - Rubin, H. AU - Coppel, R. L. AU - Mizrahi, V. PY - 2005 TI - Function of the cytochrome bc1-aa3 branch of the respiratory network in mycobacteria and network adaptation occurring in response to its disruption SP - 6300-8 N1 - Sep JF - J Bacteriol JO - Journal of bacteriology VL - 187 IS - 18 SN - 0021-9193 (Print) N1 - Function of the cytochrome bc1-aa3 branch of the respiratory network in mycobacteria and network adaptation occurring in response to its disruption N1 - 16159762 N1 - Ai43420/ai/niaid Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Electron Transport *Electron Transport Chain Complex Proteins Electron Transport Complex III/chemistry/genetics/metabolism/*physiology Electron Transport Complex IV/chemistry/metabolism Mycobacterium smegmatis/genetics/metabolism/*physiology Oxygen/metabolism Oxygen Consumption N2 - The aerobic electron transport chain in Mycobacterium smegmatis can terminate in one of three possible terminal oxidase complexes. The structure and function of the electron transport pathway leading from the menaquinol-menaquinone pool to the cytochrome bc1 complex and terminating in the aa3-type cytochrome c oxidase was characterized. M. smegmatis strains with mutations in the bc1 complex and in subunit II of cyctochome c oxidase were found to be profoundly growth impaired, confirming the importance of this respiratory pathway for mycobacterial growth under aerobic conditions. Disruption of this pathway resulted in an adaptation of the respiratory network that is characterized by a marked up-regulation of cydAB, which encodes the bioenergetically less efficient and microaerobically induced cytochrome bd-type menaquinol oxidase that is required for the growth of M. smegmatis under O2-limiting conditions. Further insights into the adaptation of this organism to rerouting of the electron flux through the branch terminating in the bd-type oxidase were revealed by expression profiling of the bc1-deficient mutant strain using a partial-genome microarray of M. smegmatis that is enriched in essential genes. Although the expression profile was indicative of an increase in the reduced state of the respiratory chain, blockage of the bc1-aa3 pathway did not induce the sentinel genes of M. smegmatis that are induced by oxygen starvation and are regulated by the DosR two-component regulator. AD - MRC/NHLS/WITS Molecular Mycobacteriology Research Unit, DST/NRF/ Centre of Excellence in Biomedical TB Research, School of Pathology, National Health Laboratory Service, Johannesburg, South Africa. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16159762 ID - 22 ER - TY - JOUR AU - Marland, Z. AU - Beddoe, T. AU - Zaker-Tabrizi, L. AU - Coppel, R. L. AU - Crellin, P. K. AU - Rossjohn, J. PY - 2005 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of an essential lipoprotein implicated in cell-wall biosynthesis in Mycobacteria SP - 1081-3 N1 - Dec 1 JF - Acta Crystallograph Sect F Struct Biol Cryst Commun JO - Acta crystallographica VL - 61 IS - Pt 12 SN - 1744-3091 (Electronic) N1 - Expression, purification, crystallization and preliminary X-ray diffraction analysis of an essential lipoprotein implicated in cell-wall biosynthesis in Mycobacteria N1 - 16511240 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Bacterial Proteins/*chemistry Cell Wall/metabolism Cloning, Molecular Crystallography, X-Ray Electrophoresis, Polyacrylamide Gel Lipopolysaccharides/chemistry Lipoproteins/*chemistry Mannosides/chemistry Mycobacterium tuberculosis/*metabolism Phosphatidylinositols/chemistry Polyethylene Glycols/chemistry Protein Structure, Tertiary X-Ray Diffraction N2 - Mycobacterium tuberculosis is a renewed cause of devastation in the developing world. Critical to the success of this re-emerging pathogen is its unusual waxy cell wall, which is rich in rare components including lipoarabinomannan (LAM) and its precursors, the phosphatidylinositol mannosides (PIMs). Balanced synthesis of these related glycolipids is intrinsic to both cell-wall integrity and virulence in M. tuberculosis and presents a promising, albeit poorly defined, therapeutic target. Here, the expression, purification and crystallization of an essential 600-amino-acid lipoprotein, LpqW, implicated in this process are reported. Crystals of LpqW were grown using 20-24%(w/v) PEG 4000, 8-16%(v/v) 2-propanol, 100 mM sodium citrate pH 5.5 and 10 mM DTT. A complete data set was collected at 2.4 A using synchrotron radiation on a crystal belonging to space group C222, with unit-cell parameters a = 188.57, b = 312.04, c = 104.15 A. Structure determination is under way. AD - Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16511240 ID - 18 ER - TY - JOUR AU - Mao, T. K. AU - Lian, Z. X. AU - Selmi, C. AU - Ichiki, Y. AU - Ashwood, P. AU - Ansari, A. A. AU - Coppel, R. L. AU - Shimoda, S. AU - Ishibashi, H. AU - Gershwin, M. E. PY - 2005 TI - Altered monocyte responses to defined TLR ligands in patients with primary biliary cirrhosis SP - 802-8 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 42 IS - 4 SN - 0270-9139 (Print) N1 - Altered monocyte responses to defined TLR ligands in patients with primary biliary cirrhosis N1 - 16175622 N1 - In Vitro Journal Article United States N1 - eng KW - Adult Aged Antigens, CD14/metabolism Cytokines/metabolism Female Flagellin/metabolism/pharmacology Humans Ligands Lipopolysaccharides/metabolism/pharmacology Liver Cirrhosis, Biliary/*immunology/*metabolism Male Membrane Glycoproteins/*metabolism Middle Aged Monocytes/drug effects/*metabolism Oligodeoxyribonucleotides/metabolism/pharmacology Poly I-C/metabolism/pharmacology Receptors, Cell Surface/*metabolism Teichoic Acids/metabolism/pharmacology Toll-Like Receptor 2 Toll-Like Receptor 3 Toll-Like Receptor 4 Toll-Like Receptor 5 Toll-Like Receptor 9 Toll-Like Receptors N2 - The role of the adaptive immune response, with regard to the development of autoantibodies, has been extensively studied in primary biliary cirrhosis (PBC). However, the importance of innate immunity has been noted only recently. Based on the proposed role of microorganisms in the pathogenesis of the disease, we hypothesize that patients with PBC possess a hyper-responsive innate immune system to pathogen-associated stimuli that may facilitate the loss of tolerance. To address this issue, we isolated peripheral blood monocytes from 33 patients with PBC and 26 age-matched healthy controls and stimulated such cells in vitro with defined ligands for toll-like receptor (TLR) 2 (lipoteichoic acid; LTA), TLR3 (polyIC), TLR4 (lipopolysaccharide; LPS), TLR5 (flagellin), and TLR9 (CpG-B). Supernatant fluids from the cultures were analyzed for levels of 5 different pro-inflammatory cytokines, interleukin (IL)-1beta, IL-6, IL-8, IL-12p70, and TNF-alpha. After in vitro challenge with TLR ligands, PBC monocytes produced higher relative levels of pro-inflammatory cytokines, particularly IL-1beta, IL-6, IL-8, and TNF-alpha, compared with controls. In conclusion, monocytes from patients with PBC appear more sensitive to signaling via select TLRs, resulting in secretion of selective pro-inflammatory cytokines integral to the inflammatory response that may be critical in the breakdown of self-tolerance. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16175622 ID - 21 ER - TY - JOUR AU - Leung, P. S. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2005 TI - Etiology of primary biliary cirrhosis: the search for the culprit SP - 327-36 N1 - Aug JF - Semin Liver Dis JO - Seminars in liver disease VL - 25 IS - 3 SN - 0272-8087 (Print) N1 - Etiology of primary biliary cirrhosis: the search for the culprit N1 - 16143948 N1 - Dk037003/dk/niddk Dk39588/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Antibody Formation Bile Ducts, Intrahepatic/pathology Epithelial Cells/physiology Genetic Predisposition to Disease Humans Ketone Oxidoreductases/metabolism Liver Cirrhosis, Biliary/*etiology/*immunology/physiopathology Mitochondria/immunology/metabolism Mitochondrial Proteins/metabolism Multienzyme Complexes Risk Factors Xenobiotics/poisoning N2 - Primary biliary cirrhosis (PBC) is an organ-specific autoimmune disease characterized by the presence of high titer antimitochondrial autoantibodies (AMAs) and destruction of intrahepatic small bile ducts. Despite vigorous efforts in the characterization of autoantibodies and bile duct histopathology, the etiology of this disease is unclear. Although there is no correlation between the titer of AMAs and disease severity, the presence of AMAs usually occurs before symptoms of liver abnormalities. We believe that the production of AMAs is not an epiphenomenon, and an understanding of the mechanism of AMA induction will shed light on the etiology of PBC. Recent studies have suggested that the induction of PBC is multifactorial, in which the primary player involves the xenobiotics modification of mitochondrial proteins or exposure to xenobiotic-modified bacterial mitochondrial protein homologs, leading to breaking of tolerance to the human mitochondrial autoantigens and eventually liver pathology in genetic susceptible individuals. We discuss the immunophysiological characteristics of biliary epithelial cells, biochemistry of the 2-oxo-acid dehydrogenase complex, and environmental and genetic factors relevant to PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16143948 ID - 24 ER - TY - JOUR AU - Kimura, Y. AU - Selmi, C. AU - Leung, P. S. AU - Mao, T. K. AU - Schauer, J. AU - Watnik, M. AU - Kuriyama, S. AU - Nishioka, M. AU - Ansari, A. A. AU - Coppel, R. L. AU - Invernizzi, P. AU - Podda, M. AU - Gershwin, M. E. PY - 2005 TI - Genetic polymorphisms influencing xenobiotic metabolism and transport in patients with primary biliary cirrhosis SP - 55-63 N1 - Jan JF - Hepatology JO - Hepatology (Baltimore, Md VL - 41 IS - 1 SN - 0270-9139 (Print) N1 - Genetic polymorphisms influencing xenobiotic metabolism and transport in patients with primary biliary cirrhosis N1 - 15690482 N1 - 39588/phs Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Aged Alleles Biological Transport/genetics Case-Control Studies Cytochrome P-450 CYP2D6/genetics Cytochrome P-450 CYP2E1/genetics Female Gene Frequency Genes, MDR Genotype Humans Liver Cirrhosis, Biliary/*genetics/*metabolism/physiopathology Male Middle Aged *Polymorphism, Genetic Receptors, Cytoplasmic and Nuclear/genetics Receptors, Steroid/genetics Severity of Illness Index Xenobiotics/*metabolism N2 - Epidemiological data suggest that environmental factors may trigger autoimmunity in genetically susceptible individuals. In primary biliary cirrhosis (PBC), it has been postulated that halogenated xenobiotics can modify self-molecules, facilitating the breakdown of tolerance to mitochondrial antigens. The transport and metabolism of xenobiotics is highly dependent on key genetic polymorphisms that alter enzymatic phenotype. We analyzed genomic DNA from 169 patients with PBC and 225 geographically and sex-matched healthy subjects for polymorphisms of genes coding for cytochromes P450 (CYPs) 2D6 (CYP2D6*4, CYP2D6*3, CYP2D6*5, and CYP2D6*6) and 2E1 (cl/c2), multidrug resistance 1 (MDR1 C3435T) P-glycoprotein, and pregnane X receptor (PXR C-25385T, C8055T, and A7635G). We compared the genotype frequencies in patients and controls and also correlated polymorphisms with PBC severity. The distributions of the studied genotypes did not significantly differ between patients and controls. However, when clinical characteristics of patients with PBC were compared according to genotype, the CYP2E1 c2 allele was associated with signs of more severe disease. In conclusion, genetic polymorphisms of CYP 2D6 and 2E1, PXR, and MDR1 do not appear to play a role in the onset of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15690482 ID - 34 ER - TY - JOUR AU - Kikuchi, K. AU - Lian, Z. X. AU - Yang, G. X. AU - Ansari, A. A. AU - Ikehara, S. AU - Kaplan, M. AU - Miyakawa, H. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2005 TI - Bacterial CpG induces hyper-IgM production in CD27(+) memory B cells in primary biliary cirrhosis SP - 304-12 N1 - Feb JF - Gastroenterology JO - Gastroenterology VL - 128 IS - 2 SN - 0016-5085 (Print) N1 - Bacterial CpG induces hyper-IgM production in CD27(+) memory B cells in primary biliary cirrhosis N1 - 15685542 N1 - Dk39588/dk/niddk Dk92310/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Antigens, CD/blood/*immunology Antigens, CD27/*blood Autoantibodies/blood B-Lymphocytes/*immunology Dinucleoside Phosphates/*pharmacology Humans Immunoglobulin M/*blood Immunologic Memory Liver Cirrhosis, Biliary/blood/*immunology Membrane Glycoproteins/blood Middle Aged Mitochondria/immunology Receptors, Cell Surface/blood Reference Values Toll-Like Receptor 9 Toll-Like Receptors N2 - BACKGROUND AND AIMS: Sera from patients with primary biliary cirrhosis (PBC) are characterized by the presence of antimitochondrial antibodies and elevated levels of immunoglobulin (Ig) M. We hypothesized that the increase in serum IgM is the result of chronic B-cell activation induced via the Toll-like receptor (TLR) signaling pathway. METHODS: We analyzed peripheral blood mononuclear cells (PBMCs) from patients with PBC and controls following incubation with CpG, a natural ligand for TLR9, and determined the basal and stimulated levels of intracellular IgM, the density of TLR9, and the contribution of specific B-cell subpopulations. RESULTS: Our data demonstrate uniquely that in vitro incubation of PBMCs from PBC with CpG-B, but not CpG-A, led to a markedly high frequency of intracellular IgM-positive B cells, associated with high levels of synthesized IgM and identified to be a function of CD27(+) memory B cells. This memory B-cell subset also expressed higher densities of TLR9 as compared with naive B cells. These results were not due to increased proliferation, as defined by 5-carboxyfluoresein diacetate succinimidyl ester labeling, or an increase in the life span of B cells, as defined by Bcl-2 expression. CONCLUSIONS: These findings for the first time identify a major role for innate immune mechanisms in the induction and persistence of abnormal humoral immune responses in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15685542 ID - 35 ER - TY - JOUR AU - Kikuchi, K. AU - Lian, Z. X. AU - Kimura, Y. AU - Selmi, C. AU - Yang, G. X. AU - Gordon, S. C. AU - Invernizzi, P. AU - Podda, M. AU - Coppel, R. L. AU - Ansari, A. A. AU - Ikehara, S. AU - Miyakawa, H. AU - Gershwin, M. E. PY - 2005 TI - Genetic polymorphisms of toll-like receptor 9 influence the immune response to CpG and contribute to hyper-IgM in primary biliary cirrhosis SP - 347-52 N1 - Jun JF - J Autoimmun JO - Journal of autoimmunity VL - 24 IS - 4 SN - 0896-8411 (Print) N1 - Genetic polymorphisms of toll-like receptor 9 influence the immune response to CpG and contribute to hyper-IgM in primary biliary cirrhosis N1 - 15878652 N1 - Journal Article England N1 - eng KW - Adult Aged Autoantibodies/*immunology Cells, Cultured Female GC Rich Sequence/immunology Humans Immunoglobulin M/*immunology Leukocytes, Mononuclear/immunology Liver Cirrhosis, Biliary/*genetics/immunology Male Membrane Glycoproteins/*genetics/immunology Middle Aged Mitochondria/immunology Oligodeoxyribonucleotides/immunology/*pharmacology *Polymorphism, Single Nucleotide/immunology Receptors, Cell Surface/*genetics/immunology Toll-Like Receptor 9 Toll-Like Receptors N2 - The serum hallmark of primary biliary cirrhosis (PBC) is the presence of anti-mitochondrial antibodies (AMA), found in 95% of patients. However, nearly every patient with PBC, including those who are AMA-negative, has an elevation in serum IgM. This hyper-IgM is neither representative of other Ig isoforms, nor is due to the levels of AMA. In fact, we have recently reported that the hyper-IgM is an innate immune response and can be induced with CpG-B with concurrent up-regulation of toll-like receptor 9 (TLR9). Based on these observations, we performed a two-tier study. First, we quantitated TLR9 genotypes in patients with PBC and controls and correlated these data with the B cell response to CpG-B. Second, based on these data, we performed an extensive TLR9 genotyping in a large cohort of patients and controls. We report herein that the 2848 AA TLR9 genotype is associated with enhanced gene expression and higher frequency of intracellular IgM(+) B cells following CpG stimulation. Interestingly, however, despite the functional association, there is no difference in the distribution of TLR9 genotypes between patients and controls. Our data emphasize the importance of dissecting the innate immune response in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Genome and Biomedical Sciences Facility, 451 E Health Sciences Drive, suite 6510, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15878652 ID - 30 ER - TY - JOUR AU - Coppel, R. L. AU - Black, C. G. PY - 2005 TI - Parasite genomes SP - 465-79 N1 - Apr 30 JF - Int J Parasitol JO - International journal for parasitology VL - 35 IS - 5 SN - 0020-7519 (Print) N1 - Parasite genomes N1 - 15826640 N1 - Dk-32094/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review England N1 - eng KW - Animals Computational Biology Disease Vectors *Genome, Protozoan Genomics Parasites/*genetics Proteomics N2 - The availability of genome sequences and the associated transcriptome and proteome mapping projects has revolutionised research in the field of parasitology. As more parasite species are sequenced, comparative and phylogenetic comparisons are improving the quality of gene prediction and annotation. Genome sequences of parasites are also providing important data sets for understanding parasite biology and identifying new vaccine candidates and drug targets. We review some of the preliminary conclusions from examination of parasite genome sequences and discuss some of the bioinformatics approaches taken in this analysis. AD - Department of Microbiology and the Victorian Bioinformatics Consortium, Monash University, Melbourne, Vic. 3800, Australia. ross.coppel@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15826640 ID - 33 ER - TY - JOUR AU - Cooke, B. M. AU - Mohandas, N. AU - Cowman, A. F. AU - Coppel, R. L. PY - 2005 TI - Cellular adhesive phenomena in apicomplexan parasites of red blood cells SP - 273-95 N1 - Sep 30 JF - Vet Parasitol JO - Veterinary parasitology VL - 132 IS - 3-4 SN - 0304-4017 (Print) N1 - Cellular adhesive phenomena in apicomplexan parasites of red blood cells N1 - 16087297 N1 - Ai44008/ai/niaid Dk32094/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. Review Netherlands N1 - eng KW - Animals Babesia bovis/pathogenicity Babesiosis/*parasitology Cattle Cattle Diseases/*parasitology Cell Adhesion/*physiology *Erythrocytes/parasitology/physiology Humans Malaria, Falciparum/*parasitology Plasmodium falciparum/pathogenicity N2 - The apicomplexan parasites Babesia and Plasmodium are related, yet phylogenetically distinct haemoprotozoa that infect red blood cells and cause severe diseases of major human and veterinary importance. A variety of cellular and molecular interactions are pivotal in many aspects of the pathogenicity of these two parasites. Comparison of the cellular and molecular mechanisms that culminate in accumulation of parasitised red blood cells in the microvasculature of cattle infected with Babesia bovis (babesiosis) and humans infected with Plasmodium falciparum (falciparum malaria) is particularly instructive given the striking similarities in the pathophysiology of these two important medical and veterinary diseases. While such adhesive phenomena have been studied extensively in malaria, they have received relatively little attention in babesiosis. In this review, we summarise the findings of more than 25 years of research into cellular adhesive phenomena in malaria and speculate on how this body of work can now be applied to Babesia parasites. Such information is fundamental if we are to learn more about the biology of Babesia parasites, the cellular and molecular mechanisms by which they cause infection and disease and how to develop novel therapeutic strategies or vaccines for both Babesia and malaria infections. AD - Department of Microbiology, Monash University, Vic. 3800, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16087297 ID - 27 ER - TY - JOUR AU - Chuang, Y. H. AU - Lian, Z. X. AU - Cheng, C. M. AU - Lan, R. Y. AU - Yang, G. X. AU - Moritoki, Y. AU - Chiang, B. L. AU - Ansari, A. A. AU - Tsuneyama, K. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2005 TI - Increased levels of chemokine receptor CXCR3 and chemokines IP-10 and MIG in patients with primary biliary cirrhosis and their first degree relatives SP - 126-32 N1 - Sep JF - J Autoimmun JO - Journal of autoimmunity VL - 25 IS - 2 SN - 0896-8411 (Print) N1 - Increased levels of chemokine receptor CXCR3 and chemokines IP-10 and MIG in patients with primary biliary cirrhosis and their first degree relatives N1 - 16243485 N1 - Dk39588/dk/niddk N01-dk-9-2310/dk/niddk Journal Article Research Support, N.I.H., Extramural England N1 - eng N2 - Infiltrating memory T cells play an important role in the destruction of the biliary tract in primary biliary cirrhosis (PBC) and inflammatory chemokines control lymphocyte traffic through their interactions with T cell chemokine receptors. In the present study, we measured plasma levels of chemokines interferon-gamma-inducible protein-10 (IP-10) and monokine induced by gamma interferon (MIG), and also studied the expression of CXCR3 chemokine receptors in 105 subjects, including 53 patients with PBC, 26 first degree relatives and 26 healthy controls. Interestingly, plasma IP-10 and MIG levels in PBC were increased significantly compared to controls and appeared to increase with disease progression. By immunohistochemistry, IP-10 and MIG expressions were evident in the portal areas in PBC. Further, the frequency of CXCR3-expressing cells in peripheral blood was also significantly higher in PBC, and CXCR3-positive cells were also found in the portal areas of diseased livers, primarily on CD4+ cells. Finally, the daughters and sisters of PBC patients also demonstrated increased plasma levels of IP-10 and MIG, but, in contrast, displayed normal frequency of CXCR3+ expressing peripheral blood lymphocytes. Our data imply a role for specific chemokine-chemokine receptor interactions in the pathogenesis of PBC and also highlight the familial risk factor. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, 451 E. Health Sciences Drive, Suite 6510, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16243485 ID - 20 ER - TY - JOUR AU - Black, C. G. AU - Wu, T. AU - Wang, L. AU - Topolska, A. E. AU - Coppel, R. L. PY - 2005 TI - MSP8 is a non-essential merozoite surface protein in Plasmodium falciparum SP - 27-35 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 144 IS - 1 SN - 0166-6851 (Print) N1 - MSP8 is a non-essential merozoite surface protein in Plasmodium falciparum N1 - 16125802 N1 - Dk-32094/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals Antigens, Protozoan/genetics/metabolism/*physiology Blotting, Western Erythrocytes/parasitology Fluorescent Antibody Technique Life Cycle Stages Plasmodium falciparum/cytology/*growth & development/metabolism Protozoan Proteins/genetics/metabolism/*physiology Transfection N2 - MSP8 is a recently identified merozoite surface protein that shares similar structural features with the leading vaccine candidate MSP1. Both proteins contain two C-terminal epidermal growth factor (EGF)-like domains, a glycosylphosphatidylinositol (GPI) anchor attachment sequence and undergo proteolytic processing. By double recombination, we have disrupted the MSP8 gene in P. falciparum 3D7 parasites, and confirmed integration by southern hybridisation and PCR. Western blot analysis of lysates from asynchronous cultures and isolated merozoites demonstrated the absence of MSP8 in two cloned knockout lines. There was no significant difference in growth rate observed between 3D7 and the cloned DeltaMSP8 lines. Thus, unlike MSP1, MSP8 is not required for asexual stage parasite growth and replication in vitro. Further analysis of the cloned lines showed that loss of MSP8 had no effect on the levels of expression of other merozoite surface proteins including MSP1-5, 7 and 10. Stage-specific immunoblots showed that MSP8 expression commences in late rings and extends throughout the rest of the erythrocytic life cycle in the 3D7 parent line, but is absent from all stages in the DeltaMSP8 transfectants. AD - Department of Microbiology and the Victorian Bioinformatics Consortium, Monash University, Clayton, Vic. 3800, Australia. casilda.black@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16125802 ID - 25 ER - TY - JOUR AU - Amano, K. AU - Leung, P. S. AU - Rieger, R. AU - Quan, C. AU - Wang, X. AU - Marik, J. AU - Suen, Y. F. AU - Kurth, M. J. AU - Nantz, M. H. AU - Ansari, A. A. AU - Lam, K. S. AU - Zeniya, M. AU - Matsuura, E. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2005 TI - Chemical xenobiotics and mitochondrial autoantigens in primary biliary cirrhosis: identification of antibodies against a common environmental, cosmetic, and food additive, 2-octynoic acid SP - 5874-83 N1 - May 1 JF - J Immunol VL - 174 IS - 9 SN - 0022-1767 (Print) N1 - Chemical xenobiotics and mitochondrial autoantigens in primary biliary cirrhosis: identification of antibodies against a common environmental, cosmetic, and food additive, 2-octynoic acid N1 - 15845458 N1 - Dk037003/dk/niddk Dk39588/dk/niddk Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. United States 1950) N1 - eng KW - Amino Acid Sequence Antibody Specificity Antigen-Antibody Reactions Autoantibodies/biosynthesis/blood/*metabolism Autoantigens/*immunology/metabolism *Cosmetics/adverse effects Cross Reactions Dihydrolipoyllysine-Residue Acetyltransferase *Environmental Exposure/adverse effects Enzyme-Linked Immunosorbent Assay Fatty Acids, Monounsaturated/*immunology/metabolism Food Additives/adverse effects Humans Liver Cirrhosis, Biliary/blood/*immunology Mitochondria/*immunology/metabolism Molecular Mimicry/immunology Molecular Sequence Data Protein Subunits/immunology/metabolism Pyruvate Dehydrogenase Complex/blood/immunology/metabolism Xenobiotics/*immunology/metabolism N2 - Emerging evidence has suggested environmental factors as causative agents in the pathogenesis of primary biliary cirrhosis (PBC). We have hypothesized that in PBC the lipoyl domain of the immunodominant E2 component of pyruvate dehydrogenase (PDC-E2) is replaced by a chemical xenobiotic mimic, which is sufficient to break self-tolerance. To address this hypothesis, based upon our quantitative structure-activity relationship data, a total of 107 potential xenobiotic mimics were coupled to the lysine residue of the immunodominant 15 amino acid peptide of the PDC-E2 inner lipoyl domain and spotted on microarray slides. Sera from patients with PBC (n = 47), primary sclerosing cholangitis (n = 15), and healthy volunteers (n = 20) were assayed for Ig reactivity. PBC sera were subsequently absorbed with native lipoylated PDC-E2 peptide or a xenobiotically modified PDC-E2 peptide, and the remaining reactivity analyzed. Of the 107 xenobiotics, 33 had a significantly higher IgG reactivity against PBC sera compared with control sera. In addition, 9 of those 33 compounds were more reactive than the native lipoylated peptide. Following absorption, 8 of the 9 compounds demonstrated cross-reactivity with lipoic acid. One compound, 2-octynoic acid, was unique in both its quantitative structure-activity relationship analysis and reactivity. PBC patient sera demonstrated high Ig reactivity against 2-octynoic acid-PDC-E2 peptide. Not only does 2-octynoic acid have the potential to modify PDC-E2 in vivo but importantly it was/is widely used in the environment including perfumes, lipstick, and many common food flavorings. AD - Division of Rheumatology, Allergy and Clinical Immunology, Genomic and Biomedical Sciences Facility, University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15845458 ID - 32 ER - TY - JOUR AU - Wang, L. AU - Webster, D. E. AU - Wesselingh, S. L. AU - Coppel, R. L. PY - 2004 TI - Orally delivered malaria vaccines: not too hard to swallow SP - 1585-94 N1 - Oct JF - Expert Opin Biol Ther JO - Expert opinion on biological therapy VL - 4 IS - 10 SN - 1744-7682 (Electronic) N1 - Orally delivered malaria vaccines: not too hard to swallow N1 - 15461570 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review England N1 - eng KW - Administration, Oral Animals Antigens, Protozoan/administration & dosage/immunology Endocytosis Erythrocytes/parasitology Feasibility Studies Humans ISCOMs/administration & dosage/immunology Immune Tolerance Malaria/prevention & control Malaria Vaccines/*administration & dosage/immunology/pharmacokinetics Mice Particle Size Plants, Genetically Modified Plasmodium/growth & development/immunology Salmonella Vaccination/legislation & jurisprudence Vaccines, DNA/administration & dosage/immunology/pharmacokinetics Vaccines, Synthetic/administration & dosage/immunology N2 - Vaccines offer efficient and cost-effective protection against a wide range of infectious diseases. Unfortunately, no effective vaccine is yet available against malaria, and this infection remains one of the most important causes of human morbidity and mortality in the developing world. Over the past two decades a number of candidate proteins for inclusion in a subunit vaccine have been identified. Malariologists believe that an effective malaria vaccine will need to include multiple proteins that induce protective immune responses against different stages of the Plasmodium life cycle. The construction of such multivalent vaccines is beset by considerable logistical difficulties, not least of which is how to deliver them to a population living in endemic areas. Compared with other routes of vaccine administration, oral delivery has several advantages that make it an attractive strategy for vaccine development. This review summarises the progress towards an oral vaccine delivery system for malaria and discusses the feasibility of this approach. AD - Monash University, Department of Microbiology, Clayton, Victoria 3800, Australia. lina.wang@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15461570 ID - 41 ER - TY - JOUR AU - Wang, L. AU - Goschnick, M. W. AU - Coppel, R. L. PY - 2004 TI - Oral immunization with a combination of Plasmodium yoelii merozoite surface proteins 1 and 4/5 enhances protection against lethal malaria challenge SP - 6172-5 N1 - Oct JF - Infect Immun JO - Infection and immunity VL - 72 IS - 10 SN - 0019-9567 (Print) N1 - Oral immunization with a combination of Plasmodium yoelii merozoite surface proteins 1 and 4/5 enhances protection against lethal malaria challenge N1 - 15385527 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Administration, Oral Animals Antibodies, Protozoan/analysis/immunology Antigens, Protozoan/genetics/*immunology Drug Therapy, Combination Escherichia coli/genetics Immunoglobulin Isotypes/analysis/immunology Malaria/blood/immunology/prevention & control Malaria Vaccines/*administration & dosage/genetics/*immunology Membrane Proteins/genetics/*immunology Merozoite Surface Protein 1/genetics/*immunology Mice Parasitemia/blood/immunology Plasmodium yoelii/genetics/*immunology Protozoan Proteins/genetics/*immunology Survival Rate N2 - Oral immunization of mice with Escherichia coli-expressed Plasmodium yoelii merozoite surface protein 4/5 or the C-terminal 19-kDa fragment of merozoite surface protein 1 induced systemic antibody responses and protected mice against lethal malaria infection. A combination of these two proteins administered orally conferred improved protection compared to that conferred by either protein administered alone. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15385527 ID - 42 ER - TY - JOUR AU - Tsuji, K. AU - Watanabe, Y. AU - Kouno, H. AU - Aisaka, Y. AU - Masanaga, T. AU - Ishihara, H. AU - Kamiyasu, M. AU - Nakanishi, T. AU - Chayama, K. AU - Asahara, T. AU - Coppel, R. AU - Gershwin, M. E. PY - 2004 TI - A case of latent primary biliary cirrhosis SP - 166-169 N1 - Mar JF - Hepatol Res VL - 28 IS - 3 SN - 1386-6346 (Print) N1 - A case of latent primary biliary cirrhosis N1 - 15036074 N1 - Journal article N1 - Eng N2 - A 53-year-old housewife was the donor when living-related donor liver transplantation (LRLT) was performed in her younger sister (49-year-old) with terminal primary biliary cirrhosis (PBC). The donor's liver histology was diagnostic and compatible with PBC, although she was negative for antimitochondrial antibody (AMA: a specific marker of PBC) by immunofluorescence and had normal liver function tests as well as no symptoms of liver disease. In this patient with latent PBC, AMA was eventually detected by immunoblotting, although it was not detected by ELISA. These findings indicate that a family history of PBC is a risk factor for the development of this disease. Our patient was diagnosed before the advent of any clinical or biochemical indicators and before or at the onset of AMA positivity, so her liver histology revealed the earliest stage of PBC. AD - Programs for Biomedical Research, Division of Frontier Medical Science, Department of Medicine and Molecular Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15036074 ID - 51 ER - TY - JOUR AU - Topolska, A. E. AU - Richie, T. L. AU - Nhan, D. H. AU - Coppel, R. L. PY - 2004 TI - Associations between responses to the rhoptry-associated membrane antigen of Plasmodium falciparum and immunity to malaria infection SP - 3325-30 N1 - Jun JF - Infect Immun JO - Infection and immunity VL - 72 IS - 6 SN - 0019-9567 (Print) N1 - Associations between responses to the rhoptry-associated membrane antigen of Plasmodium falciparum and immunity to malaria infection N1 - 15155636 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Animals Antibodies, Protozoan/*blood Antigens, Protozoan/*immunology Epitope Mapping Erythrocyte Membrane Humans Immunoglobulin G/blood Malaria, Falciparum/*epidemiology/immunology/parasitology Plasmodium falciparum/*immunology Prevalence Protozoan Proteins/*immunology Vietnam/epidemiology N2 - Rhoptry proteins participate in the invasion of red blood cells by merozoites during the malaria parasite's asexual-stage cycle. Interference with the rhoptry protein function has been shown to prevent invasion, and three rhoptry proteins have been suggested as potential components of a vaccine against malaria. Rhoptry-associated membrane antigen (RAMA) is a 170-kDa protein of Plasmodium falciparum which is processed to a 60-kDa mature form in the rhoptries. p60/RAMA is discharged from rhoptries of free merozoites and binds to the red-cell membrane before being internalized to form part of the parasitophorous vacuole of the newly developing ring. We examined the range of anti-RAMA responses in individuals living in an area of endemicity for malaria and determined its association with clinical immunity. RAMA is immunogenic during infections, and at least three epitopes within RAMA are recognized by hyperimmune sera in immunoblots. Sera from individuals living in a region of Vietnam where malaria is endemic possessed strong antibody responses toward two C-terminal regions of RAMA. Cytophilic antibody isotypes (immunoglobulin G1 [IgG1] and IgG3) predominated in humoral responses to both C-terminal epitopes. Acute episodes of P. falciparum infection result in significant boosting of levels of antibody to an epitope at the extreme C terminus of RAMA that harbors the red-cell-binding domain. Immunity to P. falciparum infection was linked to elevated levels of IgG3 responses to this functional domain of RAMA, suggesting that the region may contain a protective epitope useful for inclusion in a multiepitope vaccine against malaria. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15155636 ID - 46 ER - TY - JOUR AU - Topolska, A. E. AU - Lidgett, A. AU - Truman, D. AU - Fujioka, H. AU - Coppel, R. L. PY - 2004 TI - Characterization of a membrane-associated rhoptry protein of Plasmodium falciparum SP - 4648-56 N1 - Feb 6 JF - J Biol Chem JO - The Journal of biological chemistry VL - 279 IS - 6 SN - 0021-9258 (Print) N1 - Characterization of a membrane-associated rhoptry protein of Plasmodium falciparum N1 - 14613941 N1 - Dk-32094/dk/niddk In Vitro Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/chemistry/genetics/metabolism Base Sequence DNA, Protozoan/genetics Erythrocytes/parasitology Humans Intracellular Membranes/metabolism Malaria, Falciparum/parasitology Membrane Microdomains/parasitology Molecular Sequence Data Organelles/metabolism Plasmodium falciparum/genetics/*metabolism/pathogenicity Protozoan Proteins/chemistry/genetics/*metabolism Recombinant Proteins/chemistry/genetics/metabolism N2 - Invasive forms of apicomplexan parasites contain secretory organelles called rhoptries that are essential for entry into host cells. We present a detailed characterization of an unusual rhoptry protein of the human malaria parasite Plasmodium falciparum, the rhoptry-associated membrane antigen (RAMA) that appears to have roles in both rhoptry biogenesis and host cell invasion. RAMA is synthesized as a 170-kDa protein in early trophozoites, several hours before rhoptry formation and is transiently localized within the endoplasmic reticulum and Golgi within lipid-rich microdomains. Regions of the Golgi membrane containing RAMA bud to form vesicles that later mature into rhoptries in a process that is inhibitable by brefeldin A. Other rhoptry proteins such as RhopH3 and RAP1 are found in close apposition with RAMA suggesting direct protein-protein interactions. We suggest that RAMA is involved in trafficking of these proteins into rhoptries. In rhoptries, RAMA is proteolytically processed to give a 60-kDa form that is anchored in the inner face of the rhoptry membrane by means of the glycosylphosphatidylinositol anchor. The p60 RAMA form is discharged from the rhoptries of free merozoites and binds to the red blood cell membrane by its most C-terminal region. In early ring stages RAMA is found in association with the parasitophorous vacuole. AD - Department of Microbiology, Monash University, Wellington Road, Clayton 3800, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14613941 ID - 55 ER - TY - JOUR AU - Topolska, A. E. AU - Black, C. G. AU - Coppel, R. L. PY - 2004 TI - Identification and characterisation of RAMA homologues in rodent, simian and human malaria species SP - 237-41 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 138 IS - 2 SN - 0166-6851 (Print) N1 - Identification and characterisation of RAMA homologues in rodent, simian and human malaria species N1 - 15555735 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals Antigens, Protozoan/*analysis/chemistry/*genetics DNA, Protozoan/chemistry/isolation & purification Genes, Protozoan Membrane Proteins/*analysis/chemistry/*genetics Molecular Sequence Data Plasmodium/*chemistry/genetics Plasmodium berghei/chemistry/genetics Plasmodium chabaudi/chemistry/genetics Plasmodium knowlesi/chemistry/genetics Plasmodium vivax/chemistry/genetics Plasmodium yoelii/chemistry/genetics Protozoan Proteins/*analysis/chemistry/*genetics Sequence Analysis, DNA Species Specificity AD - Department of Microbiology, Monash University, PO Box 53, Clayton 3800, Vic., Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15555735 ID - 37 ER - TY - JOUR AU - Selmi, C. AU - Ross, S. R. AU - Ansari, A. A. AU - Invernizzi, P. AU - Podda, M. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2004 TI - Lack of immunological or molecular evidence for a role of mouse mammary tumor retrovirus in primary biliary cirrhosis SP - 493-501 N1 - Aug JF - Gastroenterology JO - Gastroenterology VL - 127 IS - 2 SN - 0016-5085 (Print) N1 - Lack of immunological or molecular evidence for a role of mouse mammary tumor retrovirus in primary biliary cirrhosis N1 - 15300582 N1 - Journal Article United States N1 - eng KW - Aged Antibodies, Viral/blood Antigens, Viral, Tumor/genetics Base Sequence Female Humans Liver/virology Liver Cirrhosis, Biliary/*virology Lymphocytes/virology Mammary Tumor Virus, Mouse/genetics/immunology/*isolation & purification Middle Aged Molecular Sequence Data Retroviridae Infections/complications/*diagnosis/immunology Tumor Virus Infections/complications/*diagnosis/immunology Viral Core Proteins/genetics N2 - BACKGROUND & AIMS: Recent observations, including a pilot clinical trial, have suggested that a human mouse mammary tumor virus (MMTV) causes primary biliary cirrhosis (PBC). We attempted to confirm such data. METHODS: We obtained sera from 101 patients (53 with PBC and 48 controls), fixed liver sections from 10 patients (8 PBC and 2 controls), fresh liver specimens (6 PBC and 6 controls), and fresh peripheral blood lymphocytes (PBLs) (10 PBC and 10 controls). We studied sera for reactivities against 3 different strains of MMTV virions, MMTV(C3H), MMTV(FM), and MMTV(LA), including goat polyclonal antibodies against MMTV virions, gp52, and p27 as positive controls. We stained liver specimens using polyclonal antibodies against MMTV and gp52 and further examined tissue samples and PBLs for specific MMTV genome sequences. RESULTS: By Western blot analysis, no detectable reactivity in any of the PBC sera against any of the 3 MMTV strains or MMTV gp52 or p27 was observed. However, viral proteins were recognized by our control positive polyclonal antibodies. We note that 13%-60% of PBC sera presented low reactivity against 2 proteins of approximately 57 and 74 kilodaltons. Such reactivity is related to the trace amounts of mitochondrial antigens in the virus preparations derived from murine mammary tumor tissue. No detectable immunohistochemical or molecular evidence for MMTV was found in the liver specimens or PBLs. CONCLUSIONS: We were unable to recapitulate the data on this specific retroviral etiology of PBC and suggest that such data could be the result of contamination. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15300582 ID - 44 ER - TY - JOUR AU - Selmi, C. AU - Invernizzi, P. AU - Keeffe, E. B. AU - Coppel, R. L. AU - Podda, M. AU - Rossaro, L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2004 TI - Epidemiology and pathogenesis of primary biliary cirrhosis SP - 264-71 N1 - Mar JF - J Clin Gastroenterol JO - Journal of clinical gastroenterology VL - 38 IS - 3 SN - 0192-0790 (Print) N1 - Epidemiology and pathogenesis of primary biliary cirrhosis N1 - 15128074 N1 - Journal Article Review United States N1 - eng KW - Humans Liver Cirrhosis, Biliary/*epidemiology/*etiology N2 - Primary biliary cirrhosis is an autoimmune disease of unknown etiology leading to progressive destruction of small intrahepatic bile ducts and eventually to liver cirrhosis and failure. It is characterized by female predominance (with most cases observed between the ages of 40 and 60) and serum autoantibodies to mitochondrial antigens as highly specific hallmarks. Epidemiologic data indicate a variable incidence and prevalence of the disease. A number of genetic factors have been indicated as playing a role in determining disease susceptibility or progression, although no definitive conclusion has been reached so far. However, as suggested by some epidemiologic observations, a number of environmental factors, including molecular mimicry by either microorganisms or xenobiotics, have also been proposed. A hypothesis gaining support is that environmental factors may trigger disease in genetically predisposed individuals. In this review, the available data regarding the epidemiology and pathogenesis of primary biliary cirrhosis will be described and discussed. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15128074 ID - 48 ER - TY - JOUR AU - Matsumura, S. AU - Van De Water, J. AU - Leung, P. AU - Odin, J. A. AU - Yamamoto, K. AU - Gores, G. J. AU - Mostov, K. AU - Ansari, A. A. AU - Coppel, R. L. AU - Shiratori, Y. AU - Gershwin, M. E. PY - 2004 TI - Caspase induction by IgA antimitochondrial antibody: IgA-mediated biliary injury in primary biliary cirrhosis SP - 1415-22 N1 - May JF - Hepatology JO - Hepatology (Baltimore, Md VL - 39 IS - 5 SN - 0270-9139 (Print) N1 - Caspase induction by IgA antimitochondrial antibody: IgA-mediated biliary injury in primary biliary cirrhosis N1 - 15122771 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Animals Antibody Specificity Caspases/*immunology/metabolism Cell Line Enzyme Activation/immunology Humans Immunoglobulin A/*immunology Immunoglobulin G/immunology Liver Cirrhosis, Biliary/*immunology/*metabolism Mitochondria, Liver/*immunology/metabolism N2 - Anti-mitochondrial antibodies (AMAs) have long been recognized as a serological hallmark of primary biliary cirrhosis (PBC). Although high titers of immunoglobulin (Ig)A AMAs are found in bile, saliva, and urine of patients, a pathogenic role for this antibody has remained elusive. Functional studies of this IgA in general have been impeded by low quantities of antibody and the inability to recover antigen-specific IgA in dimeric form. Using a newly defined synthetic group A. Streptococcus derived peptide, we purified large quantities of dimeric and monomeric IgA from patient sera. The purified IgA was incubated with Madine-Darby canine kidney (MDCK) cells transfected with the human polymeric Ig receptor (pIgR) and the cells studied by flow cytometric analysis for binding of carboxyfluorescein conjugated VAD-fmk peptide to activated caspase enzymes. A total of 87% of PBC patients that were anti-PDC-E2 positive had serum IgA that increased caspase activation in MDCK-pIgR+ cells compared to serum-derived IgA from controls with a maximum reaction 48 hours after addition of IgA. The titer of anti-PDC-E2 IgA among the PBC patients strongly correlated with caspase activation (cc = 0.88). Pre-absorption of the IgA using recombinant 2-oxo-acid dehydrogenase complex significantly diminished this activation. IgG from the same PBC patients did not induce caspase activation. These data suggest that during transcytosis through pIgR-positive cells, exposure to PDC-E2-specific dimeric IgA results in the initiation of caspase activation. In conclusion, we propose that due to an even greater concentration of dimeric IgA in biliary and mucosal secretions, constant transcytosis would render the exposed cells more susceptible to apoptosis resulting in subsequent bile duct damage. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15122771 ID - 49 ER - TY - JOUR AU - Mao, T. K. AU - Davis, P. A. AU - Odin, J. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2004 TI - Sidechain biology and the immunogenicity of PDC-E2, the major autoantigen of primary biliary cirrhosis SP - 1241-8 N1 - Dec JF - Hepatology JO - Hepatology (Baltimore, Md VL - 40 IS - 6 SN - 0270-9139 (Print) N1 - Sidechain biology and the immunogenicity of PDC-E2, the major autoantigen of primary biliary cirrhosis N1 - 15558739 N1 - Dk39588/dk/niddk Dk56839/dk/niddk Dk67003/dk/niddk Dk92310/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Amino Acid Sequence Animals Autoantigens/chemistry/*immunology/metabolism Dihydrolipoyllysine-Residue Acetyltransferase Humans Liver Cirrhosis, Biliary/*immunology/metabolism Pyruvate Dehydrogenase Complex/chemistry/*immunology/metabolism N2 - The E2 component of mitochondrial pyruvate dehydrogenase complex (PDC-E2) is the immunodominant autoantigen of primary biliary cirrhosis. Whereas lipoylation of PDC-E2 is essential for enzymatic activity and predominates under normal conditions, other biochemical systems exist that also target the lysine residue, including acylation of fatty acids or xenobiotics and ubiquitinylation. More importantly, the immunogenicity can be affected by derivatization of the lysine residue, as the recognition of lipoylated PDC-E2 by patient autoantibodies is enhanced compared with octanoylated PDC-E2. Furthermore, our laboratory has shown that various xenobiotic modifications of a peptide representing the immunodominant region of PDC-E2 are immunoreactive against patient sera. The only purported regulatory system that prevents the accumulation of potentially autoreactive PDC-E2 is glutathionylation, in which the lysine-lipoic acid moiety is further modified with glutathione during apoptosis. Interestingly, this system is found in several cell lines, including HeLa, Jurkat, and Caco-2 cells, but not in cholangiocytes and salivary gland epithelial cells, both of which are targets for destruction in primary biliary cirrhosis. Hence, the failure of this or other regulatory system(s) may overwhelm the immune system with immunogenic PDC-E2 that can initiate the breakdown of tolerance in a genetically susceptible individual. In this review the authors survey the data available on the biochemical life of PDC-E2, with particular emphasis on the lysine residue and its known interactions with machinery involved in various posttranslational modifications. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15558739 ID - 36 ER - TY - JOUR AU - Ichiki, Y. AU - Leung, P. S. AU - Ishibashi, H. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2004 TI - Mitochondria and autoimmunity in primary biliary cirrhosis SP - 743-53 N1 - Sep JF - Mitochondrion JO - Mitochondrion VL - 4 IS - 5-6 SN - 1567-7249 (Print) N1 - Mitochondria and autoimmunity in primary biliary cirrhosis N1 - 16120429 N1 - Journal Article Netherlands N1 - eng N2 - Primary biliary cirrhosis is an enigmatic autoimmune liver disease that predominantly affects women and is characterized by antimitochondrial antibodies and specific destruction of small bile ducts. Interestingly, patients with this disease not only have high titer antibodies to mitochondria, but also highly directed, liver-specific CD4 and CD8 cells directed at the same mitochondrial autoantigens. These mitochondrial autoantigens are all members of the 2-oxo dehydrogenase complex family and include the E2 component of pyruvate dehydrogenase as the major autoantigen. Moreover, the epitopes recognized by CD4, CD8 T cells and autoantibody, are all directed within the same region, namely the lipoyl domain of pyruvate dehydrogenase complex-E2. All cells in the body have mitochondria but there appear to be specific destruction of biliary cells. We believe that this specific destruction is secondary to a highly directed mucosal response that focuses on biliary cells because of the involvement of a polymeric immunoglobulin receptor, the presence of immunoglobulin A in mucosal secretions, and the unique apoptotic properties of biliary epithelium. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, TB 192, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16120429 ID - 26 ER - TY - JOUR AU - Haselhorst, T. AU - Wilson, J. C. AU - Thomson, R. J. AU - McAtamney, S. AU - Menting, J. G. AU - Coppel, R. L. AU - von Itzstein, M. PY - 2004 TI - Saturation transfer difference (STD) 1H-NMR experiments and in silico docking experiments to probe the binding of N-acetylneuraminic acid and derivatives to Vibrio cholerae sialidase SP - 346-53 N1 - Aug 1 JF - Proteins JO - Proteins VL - 56 IS - 2 SN - 1097-0134 (Electronic) N1 - Saturation transfer difference (STD) 1H-NMR experiments and in silico docking experiments to probe the binding of N-acetylneuraminic acid and derivatives to Vibrio cholerae sialidase N1 - 15211517 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't United States N1 - eng KW - Acetamides/metabolism Bacterial Proteins/*chemistry/metabolism/pharmacology Binding Sites Enzyme Inhibitors/chemistry/metabolism/pharmacology Hydrophobicity Molecular Structure N-Acetylneuraminic Acid/*analogs & derivatives/*chemistry/metabolism/pharmacology Neuraminidase/antagonists & inhibitors/*chemistry/metabolism *Nuclear Magnetic Resonance, Biomolecular Protein Binding Protein Conformation Solutions Structure-Activity Relationship Uronic Acids/chemistry/metabolism/pharmacology Vibrio cholerae/*enzymology N2 - Saturation transfer difference (STD) (1)H NMR experiments were used to probe the epitope binding characteristics of the sialidase [EC 3.2.1.18] from the bacterium Vibrio cholerae, the causative agent of cholera. Binding preferences were investigated for N-acetylneuraminic acid (Neu5Ac, 1), the product of the sialidase catalytic reaction, for the known sialidase inhibitor 5-acetamido-2,6-anhydro-3,5-dideoxy-D-glycero-D-galacto-non-2-enoic acid (Neu5Ac2en, 2), and for the uronic acid-based Neu5Ac2en mimetic iso-propyl 2-acetamido-2,4-dideoxy-alpha-L-threo-hex-4-enopyranosiduronic acid (3), in which the native glycerol side-chain of Neu5Ac2en is replaced with an O-iso-propyl ether. The STD experiments provided evidence, supporting previous studies, that Neu5Ac (1) binds to the sialidase as the alpha-anomer. Docking experiments using DOCK (version 4.0.1) revealed further information regarding the binding characteristics of the enzyme active site in complex with Neu5Ac2en (2) and the Neu5Ac2en mimetic (3), indicating an expected dominant interaction of the acetamide moiety with the protein. AD - Institute for Glycomics, Griffith University Gold Coast Campus, Queensland, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15211517 ID - 45 ER - TY - JOUR AU - Goschnick, M. W. AU - Black, C. G. AU - Kedzierski, L. AU - Holder, A. A. AU - Coppel, R. L. PY - 2004 TI - Merozoite surface protein 4/5 provides protection against lethal challenge with a heterologous malaria parasite strain SP - 5840-9 N1 - Oct JF - Infect Immun JO - Infection and immunity VL - 72 IS - 10 SN - 0019-9567 (Print) N1 - Merozoite surface protein 4/5 provides protection against lethal challenge with a heterologous malaria parasite strain N1 - 15385485 N1 - Journal Article Research Support, Non-U.S. Gov't United States N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan/immunology Antigens, Protozoan/chemistry/genetics/*immunology/isolation & purification Escherichia coli/genetics Female Malaria/immunology/*parasitology/*prevention & control Malaria Vaccines/chemistry/genetics/immunology Membrane Proteins/chemistry/genetics/*immunology/isolation & purification Mice Mice, Inbred BALB C Molecular Sequence Data Plasmodium/chemistry/classification/genetics/immunology Plasmodium yoelii/chemistry/classification/genetics/*immunology Polymorphism, Genetic/genetics Protozoan Proteins/chemistry/genetics/*immunology/isolation & purification Recombinant Proteins/chemistry/genetics/immunology/isolation & purification Sequence Analysis, DNA Species Specificity Survival Rate N2 - Immunization with merozoite surface protein 4/5 (MSP4/5), the murine malaria homologue of Plasmodium falciparum MSP4 and MSP5, has been shown to protect mice against challenge by parasites expressing the homologous form of the protein. The gene encoding MSP4/5 was sequenced from a number of Plasmodium yoelii isolates in order to assess the level of polymorphism in the protein. The gene was found to be highly conserved among the 13 P. yoelii isolates sequenced, even though many of the same isolates showed pronounced variability in their MSP1(19) sequences. Nonsynonymous mutations were detected only for the isolates Plasmodium yoelii nigeriensis N67 and Plasmodium yoelii killicki 193L and 194ZZ. Immunization and challenge of BALB/c mice showed that the heterologous MSP4/5 proteins were able to confer a level of protection against lethal Plasmodium yoelii yoelii YM challenge infection similar to that induced by immunization with the homologous MSP4/5 protein. To explore the limits of heterologous protection, mice were immunized with recombinant MSP4/5 protein from Plasmodium berghei ANKA and Plasmodium chabaudi adami DS and challenged with P. y. yoelii YM. Interestingly, significant protection was afforded by P. berghei ANKA MSP4/5, which shows 81% sequence identity with P. y. yoelii YM MSP4/5, but it was abolished upon reduction and alkylation. Significant protection was not observed for mice immunized with recombinant P. c. adami DS MSP4/5, which shows 55.7% sequence identity with P. y. yoelii YM MSP4/5. This study demonstrates the robustness of MSP4/5 in conferring protection against variant forms of the protein in a murine challenge system, in contrast to the situation found for other asexual-stage proteins, such as MSP1(19) and AMA1. AD - Department of Microbiology, Monash University, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15385485 ID - 43 ER - TY - JOUR AU - Coppel, R. L. AU - Roos, D. S. AU - Bozdech, Z. PY - 2004 TI - The genomics of malaria infection SP - 553-7 N1 - Dec JF - Trends Parasitol JO - Trends in parasitology VL - 20 IS - 12 SN - 1471-4922 (Print) N1 - The genomics of malaria infection N1 - 15522663 N1 - Dk 32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review England N1 - eng KW - Animals DNA, Protozoan/genetics *Genome, Protozoan Genomics/methods Humans Malaria, Falciparum/*parasitology Mice Plasmodium falciparum/*genetics/physiology Protozoan Proteins Transcription, Genetic N2 - Malaria research is now dominated by information flowing from the genome sequencing projects and the associated transcriptome- and proteome-mapping projects. As more species are sequenced, comparative and phylogenetic comparisons are improving the quality of gene finding, and are providing various approaches to the identification of genes important to parasite biology and the pathogenesis of disease. We are still in the early days of exploiting these data in a systematic way and the sheer volume of data presents daunting challenges. This article reviews the progress in using this genomic information and discusses opportunities for other approaches. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. ross.coppel@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15522663 ID - 39 ER - TY - JOUR AU - Cooke, B. M. AU - Mohandas, N. AU - Coppel, R. L. PY - 2004 TI - Malaria and the red blood cell membrane SP - 173-88 N1 - Apr JF - Semin Hematol JO - Seminars in hematology VL - 41 IS - 2 SN - 0037-1963 (Print) N1 - Malaria and the red blood cell membrane N1 - 15071793 N1 - Ai 44008/ai/niaid Dk 32094/dk/niddk Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Animals Antigens, Surface/metabolism Cell Adhesion/physiology Erythrocyte Membrane/parasitology/*physiology Erythrocytes/*parasitology/*physiology Host-Parasite Relations Humans Malaria/genetics/*metabolism/parasitology Membrane Proteins/genetics/metabolism Plasmodium/genetics/*physiology Protein Transport Protozoan Proteins/genetics/*metabolism N2 - Malaria is the most serious and widespread parasitic disease of humans and is arguably the commonest disease of red blood cells (RBCs). Malaria has exerted a powerful effect on human evolution and selection for resistance has led to the appearance and persistence of a number of inherited diseases. After parasite invasion, RBCs are progressively and dramatically modified. New structures appear inside the RBC and novel parasite proteins are exported to the erythrocyte cytoplasm and membrane skeleton. Radical biochemical, morphological, and rheological alterations manifest as increased membrane rigidity, reduced cell deformability, and greater adhesiveness for the vascular endothelium and other blood cells. Numerous protein-protein interactions between the malaria-parasite and the host RBC are important for many aspects of parasite biology and the pathogenesis of malaria. In addition, there are many other parasite proteins located within the infected red cell and at the membrane skeleton, for which no precise functional roles have yet been elucidated. Sequencing and annotation of the complete genome of Plasmodium falciparum, the production of proteomic and transcriptomic profiles of parasites, and the development of a transfection system for the asexual stage of the parasite are all recent achievements that should advance understanding of the molecular mechanisms that underlie the parasite-induced functional alterations in red cells. AD - Department of Microbiology, Monash University, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15071793 ID - 50 ER - TY - JOUR AU - Cooke, B. M. AU - Coppel, R. L. PY - 2004 TI - Blue skies or stormy weather: what lies ahead for malaria research? SP - 611-4 N1 - Dec JF - Trends Parasitol JO - Trends in parasitology VL - 20 IS - 12 SN - 1471-4922 (Print) N1 - Blue skies or stormy weather: what lies ahead for malaria research? N1 - 15522672 N1 - Journal Article Review England N1 - eng KW - Animals Disease Models, Animal Genome, Protozoan Humans Malaria, Falciparum/*parasitology Mice Plasmodium falciparum/genetics/*growth & development Plasmodium yoelii/genetics/*growth & development Research Design/*trends N2 - During the past ten years, our understanding of many aspects of the biology of malaria parasites has increased dramatically. In particular, the complete genome sequences of Plasmodium falciparum and Plasmodium yoelii, the availability of transcriptome and proteome profiles, and the establishment of transfection techniques for asexual-stage malaria parasites all represent major achievements from the past decade. Now that we are truly in the post-genomic phase of biological enquiry, this article highlights some of the opportunities and challenges that lie ahead, and speculates on what we should expect to achieve in the future. AD - Department of Microbiology, Monash University, Victoria 3800, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15522672 ID - 38 ER - TY - JOUR AU - Black, C. G. AU - Wang, L. AU - Topolska, A. E. AU - Finkelstein, D. I. AU - Horne, M. K. AU - Thomas, A. W. AU - Mohandas, N. AU - Coppel, R. L. PY - 2004 TI - Merozoite surface proteins 4 and 5 of Plasmodium knowlesi have differing cellular localisation and association with lipid rafts SP - 153-8 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 138 IS - 1 SN - 0166-6851 (Print) N1 - Merozoite surface proteins 4 and 5 of Plasmodium knowlesi have differing cellular localisation and association with lipid rafts N1 - 15500926 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/chemistry/genetics/*metabolism Membrane Microdomains/*metabolism Membrane Proteins/chemistry/genetics/*metabolism Molecular Sequence Data Plasmodium knowlesi/genetics/growth & development/*metabolism Protozoan Proteins/chemistry/genetics/*metabolism AD - Department of Microbiology and the Victorian Bioinformatics Consortium, Monash University, Clayton, Vic. 3800, Australia. casilda.black@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15500926 ID - 40 ER - TY - JOUR AU - Amano, K. AU - Leung, P. S. AU - Xu, Q. AU - Marik, J. AU - Quan, C. AU - Kurth, M. J. AU - Nantz, M. H. AU - Ansari, A. A. AU - Lam, K. S. AU - Zeniya, M. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2004 TI - Xenobiotic-induced loss of tolerance in rabbits to the mitochondrial autoantigen of primary biliary cirrhosis is reversible SP - 6444-52 N1 - May 15 JF - J Immunol VL - 172 IS - 10 SN - 0022-1767 (Print) N1 - Xenobiotic-induced loss of tolerance in rabbits to the mitochondrial autoantigen of primary biliary cirrhosis is reversible N1 - 15128836 N1 - Dk037003/dk/niddk Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Validation Studies United States 1950) N1 - eng KW - Animals Autoantibodies/biosynthesis/metabolism Autoantigens/*immunology Binding Sites, Antibody Binding, Competitive/immunology Dihydrolipoyllysine-Residue Acetyltransferase Female Humans Hydrocarbons, Brominated/administration & dosage/*immunology/metabolism Immunoglobulin G/metabolism Liver Cirrhosis, Biliary/enzymology/*immunology Mitochondria, Liver/enzymology/*immunology Oligonucleotide Array Sequence Analysis Oligopeptides/administration & dosage/immunology Pyruvate Dehydrogenase Complex/*immunology Rabbits *Self Tolerance Serum Albumin, Bovine/administration & dosage/immunology Thioctic Acid/immunology/metabolism Xenobiotics/administration & dosage/*immunology/metabolism N2 - Previous work has demonstrated that immunization of rabbits with the xenobiotic 6-bromohexanoate coupled to BSA breaks tolerance and induces autoantibodies to mitochondria in rabbits. Such immunized rabbits develop high-titer Abs to pyruvate dehydrogenase complex (PDC)-E2, the major autoantigen of primary biliary cirrhosis. In efforts to map the fine specificity of these autoantibodies, rabbits were immunized biweekly with 6-bromohexanoate-BSA and screened for reactivity using a unique xenobiotic-peptide-agarose microarray platform with an emphasis on identifying potential structures that mimic the molecular image formed by the association of lipoic acid with the immunodominant PDC-E2 peptide. Essentially, a total of 23 xenobiotics and lipoic acid were coupled to the 12-mer peptide backbones, PDC, a mutant PDC, and albumin. As expected, we succeeded in breaking tolerance using this small organic molecule coupled to BSA. However, unlike multiple experimental methods of breaking tolerance, we report in this study that, following continued immunization, the rabbits recover tolerance. With repeated immunization, the response to the rPDC-E2 protein increased with a gradual reduction in autoantibodies against the lipoic acid-peptide, i.e., the primary tolerance-breaking autoantigen. Detailed analysis of this system may provide strategies on how to restore tolerance in patients with autoimmune disease. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15128836 ID - 47 ER - TY - JOUR AU - Wu, C. T. AU - Eiserich, J. P. AU - Ansari, A. A. AU - Coppel, R. L. AU - Balasubramanian, S. AU - Bowlus, C. L. AU - Gershwin, M. E. AU - Van De Water, J. PY - 2003 TI - Myeloperoxidase-positive inflammatory cells participate in bile duct damage in primary biliary cirrhosis through nitric oxide-mediated reactions SP - 1018-25 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 38 IS - 4 SN - 0270-9139 (Print) N1 - Myeloperoxidase-positive inflammatory cells participate in bile duct damage in primary biliary cirrhosis through nitric oxide-mediated reactions N1 - 14512889 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Antigens, CD/analysis Antigens, Differentiation, Myelomonocytic/analysis Apoptosis Bile Ducts/chemistry/*pathology Epithelial Cells/chemistry Humans Liver Cirrhosis, Biliary/*pathology Macrophages/enzymology/*physiology Neutrophils/enzymology/*physiology Nitric Oxide/*physiology Nitric Oxide Synthase/analysis Nitric Oxide Synthase Type II Peroxidase/*analysis Tyrosine/*analogs & derivatives/analysis N2 - Previous studies have suggested that increased nitric oxide (NO)-mediated products are found in the livers of subjects with primary biliary cirrhosis (PBC), but the mechanisms involved remain enigmatic. We took advantage of immunohistochemistry and several unique monoclonal antibodies to study inflammatory cells responsible for the generation of NO, the enzymes responsible for NO production, the expression of 3-nitrotyrosine, and the presence of CD68(+) and/or myeloperoxidase (MPO)(+) cells. We examined a total of 113 liver specimens, including 64 with PBC, 19 with primary sclerosing cholangitis (PSC), 6 with non-A, non-B hepatitis, 6 with alcoholic liver disease, 4 with cryptogenic cirrhosis, 4 with biliary atresia, and 10 normal subjects. Twenty-two percent of PBC had elevated expression of 3-nitrotyrosine in their bile duct epithelial cells (BECs) (P =.0316). Furthermore, the BECs in PBC also demonstrated apoptotic changes. MPO-positive inflammatory cells were also noted adjacent to the basement membrane. In contrast, the liver of normal subjects showed few apoptotic changes in the bile ducts, with no evidence of MPO staining in the portal area. Furthermore, sections from livers of subjects with stage I or stage II PBC demonstrated significantly increased inflammatory cell infiltration (P =.0064) and elevated 3-nitrotyrosine expression in BECs (P =.0246) compared with stage III and IV. The presence of 3-nitrotyrosine was closely associated with infiltrating CD68- and/or MPO-positive cells. There was also a stage-associated difference in the presence of bile duct infiltrating cells and 3-nitrotyrosine in PBC with an increase dominant in early stage disease. In conclusion, NO and reactive oxygen species, collectively determined as 3-nitrotyrosine, are associated with bile duct destruction in PBC and are particularly prevalent in early stage disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California at Davis, TB 192, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14512889 ID - 58 ER - TY - JOUR AU - Wang, L. AU - Mohandas, N. AU - Thomas, A. AU - Coppel, R. L. PY - 2003 TI - Detection of detergent-resistant membranes in asexual blood-stage parasites of Plasmodium falciparum SP - 149-53 N1 - Aug 31 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 130 IS - 2 SN - 0166-6851 (Print) N1 - Detection of detergent-resistant membranes in asexual blood-stage parasites of Plasmodium falciparum N1 - 12946853 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals Cell Membrane/*chemistry/drug effects Detergents/*pharmacology Drug Resistance Glycosylphosphatidylinositols Membrane Microdomains/*chemistry/drug effects/metabolism Membrane Proteins/*analysis/isolation & purification Octoxynol/pharmacology Plasmodium falciparum/chemistry/drug effects/*growth & development/ultrastructure Protozoan Proteins/analysis/isolation & purification AD - Department of Microbiology and the Victorian Bioinformatics Consortium, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12946853 ID - 59 ER - TY - JOUR AU - Wang, L. AU - Kedzierski, L. AU - Wesselingh, S. L. AU - Coppel, R. L. PY - 2003 TI - Oral immunization with a recombinant malaria protein induces conformational antibodies and protects mice against lethal malaria SP - 2356-64 N1 - May JF - Infect Immun JO - Infection and immunity VL - 71 IS - 5 SN - 0019-9567 (Print) N1 - Oral immunization with a recombinant malaria protein induces conformational antibodies and protects mice against lethal malaria N1 - 12704105 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United States N1 - eng KW - Administration, Oral Animals Antibodies, Protozoan/*biosynthesis Antigens, Protozoan/*immunology Cholera Toxin/administration & dosage Dose-Response Relationship, Immunologic Female Immunization Malaria/*prevention & control Malaria Vaccines/*immunology Membrane Proteins/immunology Mice Mice, Inbred BALB C Plasmodium yoelii/*immunology Protozoan Proteins/*immunology Recombinant Proteins/immunology Vaccines, Synthetic/*immunology N2 - The increasing death toll from malaria, due to the decreasing effectiveness of current prophylactic and therapeutic regimens, has sparked a search for alternative methods of control, such as vaccines. Although several single proteins have shown some promise as subunit vaccines against sexual blood stages in experimental systems, it is clear that multicomponent vaccines are required. Many logistic difficulties make such an approach prohibitively expensive. In an effort to try to overcome some of these issues, we examined the possibility of oral immunization as a route for inducing host protective immunity. We report here that oral feeding of a malaria protein induced serum antibody levels similar to those induced by intraperitoneal immunization with Freund's adjuvant. Further, responses to conformational epitopes were induced. In the rodent challenge system, significant levels of protection to lethal challenge with malaria were induced in mice. The protective efficacy was highly correlated with antibody levels, which depended on the antigen dosage and required cholera toxin subunit B as an oral adjuvant. These findings offer new approaches to the development of a malaria vaccine and provide justification for the investigation of transgenic plants as a means of vaccine delivery. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12704105 ID - 66 ER - TY - JOUR AU - Wang, L. AU - Crouch, L. AU - Richie, T. L. AU - Nhan, D. H. AU - Coppel, R. L. PY - 2003 TI - Naturally acquired antibody responses to the components of the Plasmodium falciparum merozoite surface protein 1 complex SP - 403-12 N1 - Aug-Sep JF - Parasite Immunol JO - Parasite immunology VL - 25 IS - 8-9 SN - 0141-9838 (Print) N1 - Naturally acquired antibody responses to the components of the Plasmodium falciparum merozoite surface protein 1 complex N1 - 14651587 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antibodies, Protozoan/*blood Antigens, Protozoan/immunology Cloning, Molecular Escherichia coli Gene Expression Humans Immunoglobulin G/blood/classification Malaria, Falciparum/epidemiology/*immunology Membrane Proteins/genetics/*immunology Merozoite Surface Protein 1/immunology Parasitemia/immunology Plasmodium falciparum/*immunology Protozoan Proteins/genetics/*immunology Recombinant Fusion Proteins/biosynthesis/immunology/isolation & purification Vietnam/epidemiology N2 - Merozoite surface protein 6 (MSP6) and 7 (MSP7) of Plasmodium falciparum are peripheral membrane proteins whose cleaved products, MSP636, MSP722 and MSP719, are found on the merozoite surface as components of a non-covalently bound complex which also contains four polypeptides derived from merozoite surface protein 1 (MSP1). We have expressed both the precursor regions and the processed mature products of MSP6 and MSP7 in Escherichia coli and showed that these recombinant proteins react with human immune sera. In a set of sera collected from individuals living in malaria-endemic areas of Southern-central Vietnam, antibodies to the mature polypeptides of MSP636 and MSP722 were detected in 50.6 and 85.6% of the serum samples, whereas antibodies to the precursor regions of MSP6 and MSP7 were detected in only 12.1 and 42.5% of the serum samples, respectively. The predominant subclass of anti-MSP6 antibodies was IgG1, whereas the predominant subclass of anti-MSP7 antibodies was IgG3. In the same set of serum samples, the antibody responses to MSP119 are predominantly IgGI, whereas antibodies to merozoite surface protein 4 (MSP4) are mainly IgG3. This data is consistent with the proposition that, during malaria infection, variable proteins induce responses that are predominantly of the IgG3 isotype, and conserved proteins induce responses that are predominantly IgG1. The antibodies to MSP6, MSP7 and MSP119 all decreased at the time of infection, but increased during the convalescent period. No correlation was observed between the antibodies at the commencement of the study and absence of parasitaemia during surveillance in this population. AD - Department of Microbiology and the Victoria Bioinformatics Consortium, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14651587 ID - 54 ER - TY - JOUR AU - Waller, K. L. AU - Nunomura, W. AU - An, X. AU - Cooke, B. M. AU - Mohandas, N. AU - Coppel, R. L. PY - 2003 TI - Mature parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum binds to the 30-kDa domain of protein 4.1 in malaria-infected red blood cells SP - 1911-4 N1 - Sep 1 JF - Blood JO - Blood VL - 102 IS - 5 SN - 0006-4971 (Print) N1 - Mature parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum binds to the 30-kDa domain of protein 4.1 in malaria-infected red blood cells N1 - 12730097 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence Animals *Cytoskeletal Proteins Erythrocytes/metabolism/*parasitology Humans Malaria, Falciparum/*metabolism Membrane Proteins/chemistry/genetics/*metabolism Molecular Sequence Data *Neuropeptides *Plasmodium falciparum Protein Structure, Tertiary Protozoan Proteins/genetics/metabolism Recombinant Fusion Proteins/genetics N2 - The Plasmodium falciparum mature parasite-infected erythrocyte surface antigen (MESA) is exported from the parasite to the infected red blood cell (IRBC) membrane skeleton, where it binds to protein 4.1 (4.1R) via a 19-residue MESA sequence. Using purified RBC 4.1R and recombinant 4.1R fragments, we show MESA binds the 30-kDa region of RBC 4.1R, specifically to a 51-residue region encoded by exon 10 of the 4.1R gene. The 3D structure of this region reveals that the MESA binding site overlaps the region of 4.1R involved in the p55, glycophorin C, and 4.1R ternary complex. Further binding studies using p55, 4.1R, and MESA showed competition between p55 and MESA for 4.1R, implying that MESA bound at the IRBC membrane skeleton may modulate normal 4.1R and p55 interactions in vivo. Definition of minimal binding domains involved in critical protein interactions in IRBCs may aid the development of novel therapies for falciparum malaria. AD - Department of Medicine (Cardiology), Albert Einstein College of Medicine, 1300 Morris Park Ave, Bronx, NY 10461, USA. kwaller@aecom.yu.edu UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12730097 ID - 65 ER - TY - JOUR AU - Siekmann, J. H. AU - Allen, L. H. AU - Watnik, M. R. AU - Nestel, P. AU - Neumann, C. G. AU - Shoenfeld, Y. AU - Peter, J. B. AU - Patnik, M. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2003 TI - Titers of antibody to common pathogens: relation to food-based interventions in rural Kenyan schoolchildren SP - 242-9 N1 - Jan JF - Am J Clin Nutr JO - The American journal of clinical nutrition VL - 77 IS - 1 SN - 0002-9165 (Print) N1 - Titers of antibody to common pathogens: relation to food-based interventions in rural Kenyan schoolchildren N1 - 12499348 N1 - 35747/phs Clinical Trial Journal Article Randomized Controlled Trial Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Adolescent Antibodies, Bacterial/*biosynthesis Antibodies, Viral/*biosynthesis Case-Control Studies Child Child Nutrition Disorders/diet therapy/*immunology Child, Preschool *Diet Female Ferritins/blood Helicobacter pylori/*immunology/pathogenicity Humans Kenya Male Nutritional Status Rural Population N2 - BACKGROUND: Undernutrition is widely perceived to affect the development of an effective immune system. OBJECTIVE: We used a mini-analysis system to quantitate antibody titers and evaluate the sera of 200 Kenyan schoolchildren for antibodies to Helicobacter pylori [isotypes of immunoglobulins A (IgA), G (IgG), and M (IgM)], hepatitis A virus, rotavirus, tetanus toxoid (IgG), and a panel of recombinant malarial antigens (MSP1(19), MSP2, Ag512, MSP4, and MSP5). DESIGN: Children participated in a school-based feeding intervention with meat, milk, or nonanimal-source foods or in a nonintervention control group. Microvolumes (200 mL) of sera were analyzed at baseline and after 1 y. RESULTS: Nearly all children had elevated titers of antibody to H. pylori, hepatitis A virus, rotavirus, and malaria at the outset, despite a high prevalence of apparent biochemical micronutrient deficiencies and stunting, but many had titers of tetanus toxoid IgG antibodies below the protective concentration. Children with low hemoglobin had a greater proportion of elevated H. pylori IgM antibody titers at baseline, which suggests that current infection with H. pylori may be associated with anemia. Compared with the control subjects, only the group eating meat had a significant increase in H. pylori IgM antibodies during the intervention (P = 0.019). No other group comparisons with the control subjects were statistically significant. The additional finding that the sera of some children showed inadequate tetanus-protective antibodies, despite immunization, suggests that the vaccination program was suboptimal. CONCLUSIONS: A large battery of immune assays can be performed on microvolumes of sera. Furthermore, despite evidence of malnutrition, children do develop significant antibody-mediated responses to common pathogens. AD - Department of Nutrition, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12499348 ID - 68 ER - TY - JOUR AU - Selmi, C. AU - Balkwill, D. L. AU - Invernizzi, P. AU - Ansari, A. A. AU - Coppel, R. L. AU - Podda, M. AU - Leung, P. S. AU - Kenny, T. P. AU - Van De Water, J. AU - Nantz, M. H. AU - Kurth, M. J. AU - Gershwin, M. E. PY - 2003 TI - Patients with primary biliary cirrhosis react against a ubiquitous xenobiotic-metabolizing bacterium SP - 1250-7 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 38 IS - 5 SN - 0270-9139 (Print) N1 - Patients with primary biliary cirrhosis react against a ubiquitous xenobiotic-metabolizing bacterium N1 - 14578864 N1 - Dk39588/dk/niddk Es103019/es/niehs Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Alphaproteobacteria/*immunology/isolation & purification/*metabolism Amino Acid Sequence/genetics Bacterial Infections/complications Bacterial Proteins/genetics Case-Control Studies Dihydrolipoyllysine-Residue Acetyltransferase Feces/microbiology Female Humans Liver Cirrhosis, Biliary/genetics/*immunology/*microbiology Male Molecular Mimicry Molecular Sequence Data Pyruvate Dehydrogenase Complex/genetics Sequence Homology, Amino Acid Xenobiotics/*metabolism N2 - Infectious and environmental agents have been proposed as immunologic triggers for primary biliary cirrhosis (PBC). Recently, a ubiquitous organism that metabolizes organic compounds and estrogens, Novosphingobium aromaticivorans, has been defined. Importantly, 2 bacterial proteins have homology with the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Sera from 97 patients with PBC, 46 first-degree relatives, 10 spouses, and 195 controls were studied for reactivity against N. aromaticivorans and Escherichia coli. The reactivity was defined by absorption, affinity purification, and using monoclonal antibodies to PDC-E2. Stool samples from 20 patients with PBC and 34 controls were analyzed by polymerase chain reaction (PCR) for the presence of N. aromaticivorans. Sera from 100% of anti-PDC-E2 positive (77/77), 33% of anti-BCOADC E2 positive (1/3), and 12% of antimitochondrial antibody (AMA) negative patients with PBC (2/17) reacted with titers up to 10(-6) against two known lipoylated bacterial proteins (47 and 50 kd) from N. aromaticivorans, including patients with early disease. This titer was approximately 100- to 1,000-fold higher than against E. coli and verified by absorption, use of affinity-purified sera, and monoclonal antibody reagents. Moreover, 78 of 80 AMA-positive and 5 of 17 AMA-negative patients with PBC had antibodies against 3 other N. aromaticivorans proteins. In contrast, 0 of 195 control sera reacted against N. aromaticivorans. Approximately 25% of patients and controls had N. aromaticivorans in their fecal specimens. In conclusion, based on protein homology, capacity to metabolize xenobiotics as well as modulate estrogens, its presence in feces, and specific immunologic response, we propose that N. aromaticivorans is a candidate for the induction of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14578864 ID - 57 ER - TY - JOUR AU - Rainczuk, A. AU - Smooker, P. M. AU - Kedzierski, L. AU - Black, C. G. AU - Coppel, R. L. AU - Spithill, T. W. PY - 2003 TI - The protective efficacy of MSP4/5 against lethal Plasmodium chabaudi adami challenge is dependent on the type of DNA vaccine vector and vaccination protocol SP - 3030-42 N1 - Jun 20 JF - Vaccine JO - Vaccine VL - 21 IS - 21-22 SN - 0264-410X (Print) N1 - The protective efficacy of MSP4/5 against lethal Plasmodium chabaudi adami challenge is dependent on the type of DNA vaccine vector and vaccination protocol N1 - 12798647 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antibodies, Protozoan/blood Antigens, Protozoan/biosynthesis/genetics/*immunology Biolistics COS Cells Cercopithecus aethiops Female Genetic Vectors Immunization Schedule Immunoglobulin E/immunology Immunoglobulin G/immunology Malaria/*immunology/prevention & control Malaria Vaccines/genetics/*immunology Membrane Proteins/biosynthesis/genetics/*immunology Mice Mice, Inbred BALB C Plasmids Plasmodium chabaudi/genetics/*immunology Protozoan Proteins/biosynthesis/genetics/*immunology Survival Analysis Vaccines, DNA/genetics/*immunology N2 - The enhancement of immunogenicity of malarial DNA vaccines is important if they are to have practical application in protecting against blood-stage malaria. Here we describe three different DNA vaccine vector types used in conjunction with the blood-stage merozoite surface protein 4/5 (MSP4/5), the murine homologue of Plasmodium falciparum MSP4 and MSP5, in an attempt to enhance survival against lethal Plasmodium chabaudi adami DS blood-stage challenge. MSP4/5 was inserted into VR1020 (secretory), monocyte-chemotactic protein-3 (MCP-3) (chemoattractant), and cytotoxic T-lymphocyte antigen 4 (CTLA4) (lymph node targeting) vectors. Mice were immunized intradermally via gene-gun, IM injection, or boosting with recombinant MSP4/5 protein. Antibody responses after boosting were predominantly of the IgG1 and IgE isotypes, with low avidity antibodies produced in DNA primed groups. Despite antibody responses comparable to recombinant protein immunization, boosting mice primed with antigens encoded by MCP-3 and CTLA4 vectors did not enhance survival compared to vector control groups. Gene-gun vaccination using VR1020/MSP4/5 followed by recombinant MSP4/5 boosting, or gene-gun DNA vaccination alone using MCP-3/MSP4/5, resulted in enhanced survival compared to empty vector control mice. The results suggest that the enhancement of survival against lethal blood-stage malaria challenge after utilizing MSP4/5 DNA vaccination is therefore highly dependent on the route and type of vaccine vector employed. AD - Department of Biochemistry and Molecular Biology, The Cooperative Research Centre for Vaccine Technology, Clayton 3800, Australia. adam.rainczuk@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12798647 ID - 61 ER - TY - JOUR AU - Leung, P. S. AU - Quan, C. AU - Park, O. AU - Van de Water, J. AU - Kurth, M. J. AU - Nantz, M. H. AU - Ansari, A. A. AU - Coppel, R. L. AU - Lam, K. S. AU - Gershwin, M. E. PY - 2003 TI - Immunization with a xenobiotic 6-bromohexanoate bovine serum albumin conjugate induces antimitochondrial antibodies SP - 5326-32 N1 - May 15 JF - J Immunol VL - 170 IS - 10 SN - 0022-1767 (Print) N1 - Immunization with a xenobiotic 6-bromohexanoate bovine serum albumin conjugate induces antimitochondrial antibodies N1 - 12734383 N1 - Dk39588/dk/niddk Es103019/es/niehs Journal Article Research Support, U.S. Gov't, P.H.S. United States 1950) N1 - eng KW - Animals Antibody Specificity Autoantibodies/*biosynthesis/blood/physiology Autoantigens/immunology Dihydrolipoyllysine-Residue Acetyltransferase Epitopes/blood/immunology Female Haptens/administration & dosage/immunology Humans Hydrocarbons, Brominated/administration & dosage/*immunology Immunization/methods Liver Cirrhosis, Biliary/enzymology/immunology Mitochondria/*immunology/metabolism Molecular Mimicry/immunology Pyruvate Dehydrogenase Complex/analysis/antagonists & inhibitors/blood/immunology Rabbits Self Tolerance Serum Albumin, Bovine/administration & dosage/*immunology Xenobiotics/administration & dosage/*immunology N2 - The E2 subunit of pyruvate dehydrogenase complex (PDC-E2) is the major autoantigen recognized by antimitochondrial Abs (AMA) in primary biliary cirrhosis (PBC). Recently, we replaced the lipoic acid moiety of PDC-E2 with a battery of synthetic structures designed to mimic a xenobiotically modified lipoyl hapten on a 12-aa peptide that was found within the immunodominant autoepitope of PDC-E2 and demonstrated that AMA in PBC reacted against several organic modified mimotopes as well as, or sometimes significantly better than, the native lipoyl domain. Based on this data, we immunized rabbits with one such xenobiotic organic compound, 6-bromohexanoate, coupled to BSA. One hundred percent of immunized rabbits developed AMA that have each and every characteristic of human AMAs with reactivity against PDC-E2, E2 subunit of branched chain 2-oxo-acid dehydrogenase, and E2 subunit of 2-oxoglutarate dehydrogenase complex. The rabbit AMA also inhibited enzymatic function of PDC-E2 and, importantly, binds to peptide sequences not present in the xenobiotic carrier immunogen. In contrast, BSA-immunized controls did not produce such activity. Our observation that animals immunized with a xenobiotic BSA complex produce autoantibodies that react not only with the xenobiotic, but also with mitochondrial autoantigens recognized by autoimmune PBC sera, suggests that environmental xenobiotic agents can be a risk factor for the induction of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12734383 ID - 64 ER - TY - JOUR AU - Leung, P. S. AU - Park, O. AU - Matsumura, S. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2003 TI - Is there a relation between Chlamydia infection and primary biliary cirrhosis? SP - 227-33 N1 - Jun-Dec JF - Clin Dev Immunol JO - Clinical & developmental immunology VL - 10 IS - 2-4 SN - 1740-2522 (Print) N1 - Is there a relation between Chlamydia infection and primary biliary cirrhosis? N1 - 14768955 N1 - Ai 39558/ai/niaid Journal Article Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Chlamydia/genetics/immunology Chlamydia Infections/blood/*complications/*immunology/microbiology Escherichia coli/immunology Humans Immunohistochemistry Liver Cirrhosis, Biliary/*etiology/immunology/*microbiology RNA, Ribosomal, 16S/genetics N2 - Over the past two decades, a number of studies have failed to provide direct evidence of specific microbial chronic infection in primary biliary cirrhosis (PBC). However, a recent report suggests that there is a specific association of Chlamydia pneumoniae in patients with PBC and that C. pneumoniae or similar antigens might play a role in the pathogenesis of disease. To determine if Chlamydia infection is associated with PBC, we applied a combination of immunological and molecular approaches to investigate (a) the serological reactivity against two common Chlamydia human pathogens, C. pneumoniae and C. trachomatis, by immunoblotting, (b) the presence of Chlamydia in liver samples of patients with PBC and controls by PCR amplification of Chlamydia specific 16S rRNA and (c) the presence of Chlamydia proteins in liver samples of patients with PBC and controls by immunohistochemical staining. By immunoblotting, C. trachomatis and C. pneumoniae specific serological antibodies were found in 52/57 (91.2%) AMA positive PBC, 7/33 (21/2%) of AMA negative PBC, 1/25 (4%) PSC, 0/15 (0%) Sjorgen's syndrome and 0/20 (0%) systemic lupus erythematosus patients and 0/20 (0%) healthy volunteers at 1:200 sera dilution. PBC sera reacted to Chlamydia and E. coli lysates in western blots up to a maximum of 10(-4) dilution. However, PCR amplification of the Chlamydia specific 16S rRNA gene was negative in 25/25 PBC livers but positive in 1/4 PSC liver, 3/6 in other liver disease controls and 1/4 normal liver samples. While two commercially available specific monoclonal antibodies stained positive controls (Chlamydia infected HEp-2 cells) they failed to detect Chlamydia antigens in PBC livers. The detection of Chlamydia specific antibodies but not Chlamydia rRNA gene and Chlamydia antigens in PBC suggests that Chlamydia infection is not involved in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis, CA 95616, USA. psleung@ucdavis.edu UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14768955 ID - 52 ER - TY - JOUR AU - Kita, H. AU - Nalbandian, G. AU - Keeffe, E. B. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2003 TI - Pathogenesis of primary biliary cirrhosis SP - 821-39 N1 - Nov JF - Clin Liver Dis JO - Clinics in liver disease VL - 7 IS - 4 SN - 1089-3261 (Print) N1 - Pathogenesis of primary biliary cirrhosis N1 - 14598794 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Apoptosis/immunology Autoantibodies/*immunology Cytokines/immunology Epithelial Cells/immunology Humans *Immunogenetics Liver/immunology Liver Cirrhosis, Biliary/*immunology/*physiopathology Lymphocytes/immunology Mitochondria/immunology Xenobiotics/immunology N2 - Primary biliary cirrhosis is an enigmatic autoimmune disease that predominantly affects women. The serologic signatures of PBC are high titer antimitochondrial antibodies that are directed at the inner lipoyl domains of the 2-oxo-dehydrogenase enzymes, particularly PDC-E2. Of note, is that the antibody response and the CD4 and CD8 response, are all directed at a similar epitope, the inner lipoyl domain. This unique immunologic response suggests that modification of the inner lipoyl domain is associated with the immunogenetic basis of disease. AD - School of Medicine, University of California at Davis, One Shields Avenue, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14598794 ID - 56 ER - TY - JOUR AU - Kita, H. AU - Ansari, A. A. AU - He, X. S. AU - Lian, Z. X. AU - Van de Water, J. AU - Coppel, R. L. AU - Luketic, V. AU - Kaplan, M. AU - Inamori, H. AU - Isoda, N. AU - Sugano, K. AU - Imawari, M. AU - Gershwin, M. E. PY - 2003 TI - Proteasome is required for class I-restricted presentation by Fcgamma receptor-mediated endocytosis in primary biliary cirrhosis SP - 175-82 N1 - Sep JF - J Autoimmun JO - Journal of autoimmunity VL - 21 IS - 2 SN - 0896-8411 (Print) N1 - Proteasome is required for class I-restricted presentation by Fcgamma receptor-mediated endocytosis in primary biliary cirrhosis N1 - 12935787 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Acetylcysteine/*analogs & derivatives/pharmacology *Antigen Presentation Cysteine Endopeptidases/*physiology Dendritic Cells/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Endocytosis Histocompatibility Antigens Class I/metabolism Humans Liver Cirrhosis, Biliary/*enzymology/*immunology Multienzyme Complexes/antagonists & inhibitors/*physiology Peptide Fragments/genetics/immunology Proteasome Endopeptidase Complex Pyruvate Dehydrogenase Complex/genetics/*immunology Receptors, IgG/metabolism Signal Transduction T-Lymphocyte Subsets/immunology T-Lymphocytes, Cytotoxic/immunology N2 - There is a considerable database on the effector mechanisms for CD8 recognition of PDC-E2 in primary biliary cirrhosis (PBC). In particular, the specific roles of MHC class I, the mitochondrial autoepitope, and the liver-specific T cell precursor frequency, are defined for HLA-A2.1 patients. There is evidence for a role of MHC class I-mediated presentation of exogenous antigens, or cross-presentation, in the development of the antimitochondrial response and a contributory role of Fcgamma receptor-mediated uptake of autoantigen-autoantibody complexes for the induction of a PDC-E2 specific autoreactive CTL response. Based on this background, we examined potential intracellular pathways for processing the immunodominant mitochondrial autoantigen, PDC-E2, by dendritic cells (DC). In particular, we studied the effects of the proteasome inhibitor lactacystin and the endosomal acidification inhibitor bafilomycin on the induction of PDC-E2-specific CTL response in PBC. Importantly, our data indicate that pre-treatment with either lactacystin or bafilomycin inhibits the PDC-E2 immune complex-induced CTL response. The processing and presentation of PDC-E2 by CD8(+)T cells is mediated by proteasomes and facilitated by Fcgamma receptor-mediated endocytosis. This data reflects another layer of interaction between components of the immune system in the development of autoimmunity. Further characterization of autoantigen uptake and processing may lead to potential therapeutic intervention. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, TB 192, One Shields Avenue, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12935787 ID - 60 ER - TY - JOUR AU - Bruggraber, S. F. AU - Leung, P. S. AU - Amano, K. AU - Quan, C. AU - Kurth, M. J. AU - Nantz, M. H. AU - Benson, G. D. AU - Van de Water, J. AU - Luketic, V. AU - Roche, T. E. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2003 TI - Autoreactivity to lipoate and a conjugated form of lipoate in primary biliary cirrhosis SP - 1705-13 N1 - Dec JF - Gastroenterology JO - Gastroenterology VL - 125 IS - 6 SN - 0016-5085 (Print) N1 - Autoreactivity to lipoate and a conjugated form of lipoate in primary biliary cirrhosis N1 - 14724823 N1 - Dk39588/dk/niddk Es103019/es/niehs Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Animals Antibodies/*blood *Autoimmunity Epitopes Hemocyanin/immunology Humans Immunoglobulin Isotypes/blood Liver Cirrhosis, Biliary/*immunology Pyruvate Dehydrogenase Complex/immunology Rabbits Serum Albumin/immunology Thioctic Acid/*immunology N2 - BACKGROUND & AIMS: Although considerable effort has been directed toward the mapping of peptide epitopes by autoantibodies, the role of nonprotein molecules has been less well studied. The immunodominant autoantigen in primary biliary cirrhosis (PBC), E2 components of pyruvate dehydrogenase complexes (PDC-E2), has a lipoate molecule bonded to the domain to which autoantibodies are directed. METHODS: We examined sera from patients with PBC (n = 105), primary sclerosing cholangitis (n = 70), and rheumatoid arthritis (n = 28) as well as healthy volunteers (n = 43) for reactivity against lipoic acid. The lipoic acid hapten specificity of the reactive antibodies in PBC sera was determined following incubation of aliquots of the sera with human serum albumin (HSA), lipoylated HSA (HSA-LA), PDC-E2, lipoylated PDC-E2, polyethylene glycol (PEG), lipoylated PEG, free lipoic acid, and synthetic molecular mimics of lipoic acid. RESULTS: Anti-lipoic acid specific antibodies were detected in 81% (79 of 97) of antimitochondrial antibody (AMA)-positive patients with PBC but not in controls. Two previously unreported specificities in AMA-positive sera that recognize free lipoic acid and a carrier-conjugated form of lipoic acid were also identified. CONCLUSIONS: We hypothesize that conjugated form(s) of native or xenobiotic lipoic acid mimics contribute to the initiation and perpetuation of autoimmunity by at first breaking self-tolerance and participating in subsequent determinant spreading. The variability in the immunoreactive carrier/lipoate conjugates provides an experimental framework on which potential mechanisms for the breakdown of self-tolerance following exposure to xenobiotics can be investigated. The data have implications for patients taking lipoic acid as a dietary supplement. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis Medical School, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14724823 ID - 53 ER - TY - JOUR AU - Black, C. G. AU - Wang, L. AU - Wu, T. AU - Coppel, R. L. PY - 2003 TI - Apical location of a novel EGF-like domain-containing protein of Plasmodium falciparum SP - 59-68 N1 - Mar JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 127 IS - 1 SN - 0166-6851 (Print) N1 - Apical location of a novel EGF-like domain-containing protein of Plasmodium falciparum N1 - 12615336 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Base Sequence Cell Polarity Epidermal Growth Factor/*chemistry Female Humans Life Cycle Stages Malaria, Falciparum/epidemiology/parasitology Mice Mice, Inbred BALB C Molecular Sequence Data Plasmodium falciparum/*chemistry/immunology/isolation & purification Protein Structure, Tertiary Protozoan Proteins/*analysis/*chemistry/genetics Rabbits Sensitivity and Specificity N2 - Using bioinformatics analyses of the unfinished malaria genome sequence, we have identified a novel protein of Plasmodium falciparum that contains two epidermal growth factor (EGF)-like domains near the C-terminus of the protein. The sequence contains a single open reading frame of 1572bp with the potential to encode a protein of 524 residues containing hydrophobic regions at the extreme N- and C-termini which appear to represent signal peptide and glycosylphosphatidylinositol (GPI)-attachment sites, respectively. RT-PCR analysis has confirmed that the novel gene is transcribed in asexual stages of P. falciparum. Antibodies to the EGF-like domains of the novel protein are highly specific and do not cross-react with the EGF-like domains of MSP1, MSP4, MSP5 or MSP8 expressed as GST fusion proteins. Antisera to the C-terminal fragments react with two bands of 80 and 36kDa in P. falciparum parasite lysates whereas antisera to the most N-terminal fusion protein only recognises the 80kDa band, suggesting that the novel protein may undergo processing in a similar way to MSP1 and MSP8, but with fewer cleavage events. Immunoblot analysis of stage-specific parasite samples reveals that the protein is present in trophozoites, schizonts and in isolated merozoites. The protein partitions in the detergent-enriched phase after Triton X-114 fractionation and is localised to the surfaces of trophozoites, schizonts and free merozoites in an apical distribution. Based on the accepted nomenclature in the field we now designate this protein MSP10. We have shown that the MSP10 fusion proteins are in a conformation that can be recognised by human immune sera and that there is very limited sequence diversity in an approximately lkb region of MSP10, encompassing the two EGF-like domains. A sequence similar to MSP10 can be identified in the available P. yoelii genomic sequence, offering the possibility of ascertaining whether this novel protein can induce host protective responses in an in vivo model. AD - Department of Microbiology and the Victorian Bioinformatics Consortium, P.O. Box 53, Monash University, 3800 Victoria, Australia. casilda.black@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12615336 ID - 67 ER - TY - JOUR AU - Wang, L. AU - Marshall, V. M. AU - Coppel, R. L. PY - 2002 TI - Limited polymorphism of the vaccine candidate merozoite surface protein 4 of Plasmodium falciparum SP - 301-3 N1 - Apr 9 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 120 IS - 2 SN - 0166-6851 (Print) N1 - Limited polymorphism of the vaccine candidate merozoite surface protein 4 of Plasmodium falciparum N1 - 11897136 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/chemistry/*genetics/immunology Base Sequence Molecular Sequence Data Plasmodium falciparum/*genetics Polymorphism, Genetic/*genetics Protozoan Proteins/chemistry/*genetics/immunology Protozoan Vaccines/genetics/immunology Sequence Homology, Nucleic Acid AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11897136 ID - 84 ER - TY - JOUR AU - Wang, L. AU - Coppel, R. L. PY - 2002 TI - Triton X-114 phase partitioning for antigen characterization SP - 581-5 JF - Methods Mol Med JO - Methods in molecular medicine VL - 72 SN - 1543-1894 (Print) N1 - Triton X-114 phase partitioning for antigen characterization N1 - 12125157 N1 - Journal Article United States N1 - eng KW - Animals Antigens, Protozoan/*isolation & purification Detergents Erythrocytes/*parasitology Humans Membrane Proteins/isolation & purification Plasmodium/*isolation & purification *Polyethylene Glycols AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12125157 ID - 78 ER - TY - JOUR AU - Waller, K. L. AU - Nunomura, W. AU - Cooke, B. M. AU - Mohandas, N. AU - Coppel, R. L. PY - 2002 TI - Mapping the domains of the cytoadherence ligand Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) that bind to the knob-associated histidine-rich protein (KAHRP) SP - 125-9 N1 - Jan JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 119 IS - 1 SN - 0166-6851 (Print) N1 - Mapping the domains of the cytoadherence ligand Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) that bind to the knob-associated histidine-rich protein (KAHRP) N1 - 11755194 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals Binding Sites Cell Adhesion Cytoplasm/metabolism/parasitology Erythrocyte Membrane/metabolism/parasitology Kinetics Ligands Peptides/*metabolism Plasmodium falciparum/*metabolism Polymerase Chain Reaction Protein Binding Protein Structure, Tertiary Protozoan Proteins/*chemistry/genetics/*metabolism Recombinant Proteins/chemistry/genetics/metabolism Sequence Deletion/genetics Thermodynamics AD - Department of Microbiology, Monash University, Wellington Road., Vic. 3800, Clayton, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11755194 ID - 90 ER - TY - JOUR AU - Sasaki, M. AU - Long, S. A. AU - Van De Water, J. AU - He, X. S. AU - Shultz, L. AU - Coppel, R. L. AU - Ansari, A. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 2002 TI - The SJL/J mouse is not a model for PBC SP - 1284-6 N1 - May JF - Hepatology JO - Hepatology (Baltimore, Md VL - 35 IS - 5 SN - 0270-9139 (Print) N1 - The SJL/J mouse is not a model for PBC N1 - 11981783 N1 - Letter United States N1 - eng KW - Animals *Disease Models, Animal Female Liver Cirrhosis, Biliary/*pathology Mice *Mice, Inbred Strains UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11981783 ID - 82 ER - TY - JOUR AU - Sasaki, M. AU - Allina, J. AU - Odin, J. A. AU - Thung, S. N. AU - Coppel, R. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 2002 TI - Autoimmune cholangitis in the SJL/J mouse is antigen non-specific SP - 103-11 N1 - Jun JF - Dev Immunol JO - Developmental immunology VL - 9 IS - 2 SN - 1044-6672 (Print) N1 - Autoimmune cholangitis in the SJL/J mouse is antigen non-specific N1 - 12739787 N1 - Journal Article England N1 - eng KW - Adjuvants, Immunologic/administration & dosage Animals Autoantibodies/biosynthesis Autoantigens/administration & dosage Autoimmune Diseases/*etiology/immunology/pathology Bile Ducts, Intrahepatic/pathology Cholangitis/*etiology/immunology/pathology Chronic Disease Disease Models, Animal Freund's Adjuvant/administration & dosage Immunization Interferon-gamma, Recombinant/administration & dosage Liver Cirrhosis, Biliary/etiology/immunology/pathology Mice Mitochondria/enzymology/immunology Pyruvate Dehydrogenase Complex/administration & dosage/immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune disease characterized by intrahepatic bile duct destruction and the production of anti-mitochondrial antibodies (AMA). The absence of an animal model has been a striking impedance in defining the molecular basis of disease. Previous work has suggested that SJL/J mice immunize with the pyruvate dehydrogenase complex (PDC-E2), the major mitochondrial autoantigen of PBC, leads to the development of lymphoid cell infiltration in portal tracts and a model system coined autoimmune cholangitis. We hypothesized that this pathology would be augmented if immunization occurred in the presence of IFN-gamma injections. Accordingly, SJL/J mice were immunized with PDC-E2 and, for purpose of control, alpha-casein. Subgroups of mice were also treated with exogenous IFN-gamma. As expected, mice immunized with PDC-E2, with or without IFN-gamma, developed high titer AMAs. In contrast, mice immunized with alpha-casein, develop antinuclear antibodies. More importantly, the livers from mice immunized with PDC-E2 and/or those immunized with alpha-casein all displayed lymphoid cell infiltration to the portal tracts, irrespective of bile duct size. Indeed, there was no significant difference between the experimental and the control groups by histologic analysis. Thus, autoimmune cholangitis in these mice is antigen non-specific. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12739787 ID - 63 ER - TY - JOUR AU - Plebanski, M. AU - Proudfoot, O. AU - Pouniotis, D. AU - Coppel, R. L. AU - Apostolopoulos, V. AU - Flannery, G. PY - 2002 TI - Immunogenetics and the design of Plasmodium falciparum vaccines for use in malaria-endemic populations SP - 295-301 N1 - Aug JF - J Clin Invest JO - The Journal of clinical investigation VL - 110 IS - 3 SN - 0021-9738 (Print) N1 - Immunogenetics and the design of Plasmodium falciparum vaccines for use in malaria-endemic populations N1 - 12163446 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Animals Antibodies, Protozoan/immunology Antigen Presentation/immunology Antigens, CD36/immunology Cytokines/immunology Endemic Diseases Evolution Humans Immunity, Active/immunology Immunity, Natural/immunology Immunogenetics Intercellular Adhesion Molecule-1/immunology Malaria Vaccines/*immunology Malaria, Falciparum/immunology/*prevention & control Plasmodium falciparum/genetics/*immunology Receptors, Fc/immunology AD - Vaccine Development and Infectious Diseases Unit, The Austin Research Institute, A & RMC Hospital Campus, Heidelberg, Victoria, Australia. mplebans@ari.unimelb.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12163446 ID - 76 ER - TY - JOUR AU - Menting, J. G. AU - Coppel, R. L. PY - 2002 TI - Immunoprecipitation for antigen localization SP - 587-604 JF - Methods Mol Med JO - Methods in molecular medicine VL - 72 SN - 1543-1894 (Print) N1 - Immunoprecipitation for antigen localization N1 - 12125158 N1 - Journal Article United States N1 - eng KW - Animals Antibodies, Protozoan Antigens, Protozoan/*analysis/isolation & purification Chromatography, Affinity/methods Humans Immunoassay/methods Indicators and Reagents Plasmodium/growth & development/*immunology Plasmodium falciparum/growth & development/immunology Precipitin Tests/methods AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12125158 ID - 77 ER - TY - JOUR AU - Matsumura, S. AU - Van De Water, J. AU - Kita, H. AU - Coppel, R. L. AU - Tsuji, T. AU - Yamamoto, K. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2002 TI - Contribution to antimitochondrial antibody production: cleavage of pyruvate dehydrogenase complex-E2 by apoptosis-related proteases SP - 14-22 N1 - Jan JF - Hepatology JO - Hepatology (Baltimore, Md VL - 35 IS - 1 SN - 0270-9139 (Print) N1 - Contribution to antimitochondrial antibody production: cleavage of pyruvate dehydrogenase complex-E2 by apoptosis-related proteases N1 - 11786955 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Adult *Apoptosis Autoantibodies/*biosynthesis Autoantigens/immunology Caspase 3 Caspase 6 Caspase 8 Caspase 9 Caspases/metabolism Dihydrolipoyllysine-Residue Acetyltransferase Endopeptidases/*metabolism Enzyme Activation Female Granzymes Humans Immunohistochemistry Liver/enzymology Liver Cirrhosis, Biliary/*enzymology/immunology Middle Aged Mitochondria/*immunology Peptide Fragments/immunology Pyruvate Dehydrogenase Complex/immunology/*metabolism Recombinant Proteins/metabolism Serine Endopeptidases/metabolism N2 - Patients with PBC produce a directed, specific response to a single immunodominant autoepitope of PDC-E2 within the inner lipoyl domain. In contrast, immunized animals react to multiple epitopes and rarely recognize the inner lipoyl domain. In other autoimmune diseases, apoptosis plays a critical role in antigen presentation; the caspases and granzyme B are the key proteases in the generation of autoepitopes. To determine the specific cleavage pattern of full-length recombinant PDC-E2, we performed in vitro digestion with caspases-3, -6, -8 and granzyme B. The resulting fragments were immunoblotted and probed with an extensive panel of monoclonal anti-PDC-E2 antibodies and sera from patients with PBC. Interestingly, on granzyme B digestion, PDC-E2 lost reactivity, suggesting the destruction of the immunodominant epitope. Because this site contains the major epitope for both B cells and T cells, it suggests that granzyme B is unlikely to be involved in generation of autoepitopes in primary biliary cirrhosis (PBC). In contrast, following treatment with the caspase enzymes, immunoreactive fragments were generated. Indeed, by confocal microscopy, activated caspase-3 is found in the marginal hepatocytes and bile ducts. Moreover, caspase-3 staining was strongest in the small intrahepatic bile ducts, the major site of tissue destruction in PBC. In conclusion, these data suggest that following apoptosis, the caspase family of proteolytic enzymes have the potential to generate immunogenic fragments that contribute to the autoantigen reservoir and the production of antimitochondrial antibodies. These findings are also consistent with the generation of an autoimmune response against an intracellular antigen that evades catabolism during apoptosis. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11786955 ID - 87 ER - TY - JOUR AU - Matsumura, S. AU - Kita, H. AU - He, X. S. AU - Ansari, A. A. AU - Lian, Z. X. AU - Van De Water, J. AU - Yamamoto, K. AU - Tsuji, T. AU - Coppel, R. L. AU - Kaplan, M. AU - Gershwin, M. E. PY - 2002 TI - Comprehensive mapping of HLA-A0201-restricted CD8 T-cell epitopes on PDC-E2 in primary biliary cirrhosis SP - 1125-34 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 36 IS - 5 SN - 0270-9139 (Print) N1 - Comprehensive mapping of HLA-A0201-restricted CD8 T-cell epitopes on PDC-E2 in primary biliary cirrhosis N1 - 12395322 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence CD8-Positive T-Lymphocytes/cytology/*immunology/metabolism Cell Line Dihydrolipoyllysine-Residue Acetyltransferase Epitope Mapping Epitopes, T-Lymphocyte/immunology HLA-A Antigens/*genetics/immunology/metabolism Humans Liver Cirrhosis, Biliary/*genetics/*immunology Phenotype Protein Binding Pyruvate Dehydrogenase Complex/*genetics/immunology/metabolism N2 - Growing evidence has implicated the involvement of autoreactive T lymphocytes in the pathogenesis of primary biliary cirrhosis (PBC). We have recently taken advantage of motif prediction analysis of HLA-A*0201 and identified the first major histocompatibility complex (MHC) class I restricted epitope, amino acids 159 to 167 on E2 components of pyruvate dehydrogenase complexes (PDC-E2), the major mitochondrial antigens in PBC. The mechanisms involved in the selection of epitope peptide(s) that comprise the PDC-E2-specific autoreactive cytotoxic T lymphocytes (CTLs) are unknown and likely involve other epitopes on PDC-E2 restricted by MHC class I molecules. To address this issue, a comprehensive mapping of the CTL epitope repertoire on the PDC-E2 molecule that binds HLA-A*0201 was performed to provide further clues regarding the role of CTLs. We used the T2 cell line to screen 79 overlapping 15mer peptides, spanning the entire PDC-E2 molecule. Six of the 79 peptides exhibited significantly higher binding activity to HLA-A*0201 than the other 15mer peptides. Two of these 6 peptides induced CTL lines from patients with PBC. Fine mapping with N-terminus or C-terminus truncated peptides identified 10mer peptide, PDC-E2 amino acids 165 to 174, which is a novel CD8 epitope restricted by HLA-A*0201. In conclusion, using a combination of the 15mer peptide library screening with the T2 binding assay and also the induction of CTL lines with candidate peptides, we have defined a novel HLA-A*0201-restricted epitope PDC-E2 165 to 174 in patients with PBC. These data will become important in the development of altered peptide ligands to modulate disease activity. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12395322 ID - 72 ER - TY - JOUR AU - Kita, H. AU - Naidenko, O. V. AU - Kronenberg, M. AU - Ansari, A. A. AU - Rogers, P. AU - He, X. S. AU - Koning, F. AU - Mikayama, T. AU - Van De Water, J. AU - Coppel, R. L. AU - Kaplan, M. AU - Gershwin, M. E. PY - 2002 TI - Quantitation and phenotypic analysis of natural killer T cells in primary biliary cirrhosis using a human CD1d tetramer SP - 1031-43 N1 - Oct JF - Gastroenterology JO - Gastroenterology VL - 123 IS - 4 SN - 0016-5085 (Print) N1 - Quantitation and phenotypic analysis of natural killer T cells in primary biliary cirrhosis using a human CD1d tetramer N1 - 12360465 N1 - Ai45053/ai/niaid Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Antigens, CD1/*genetics Baculoviridae/genetics Flow Cytometry Galactosylceramides/immunology Humans Immunophenotyping Killer Cells, Natural/chemistry/*immunology Liver/cytology/immunology Liver Cirrhosis, Biliary/*immunology Lymph Nodes/cytology/immunology Receptors, Antigen, T-Cell, alpha-beta/analysis Recombinant Proteins/genetics/immunology N2 - BACKGROUND & AIMS: Natural killer T (NKT) cells are a subset of lymphocytes incriminated in playing an important role in the modulation of the innate immune response and the development of autoimmunity. However, there have been only limited studies attempting to quantitate the number of NKT cells in autoimmune disease, particularly because of difficulties associated with definition of this subpopulation. METHODS: We used a human CD1d (hCD1d) tetramer produced by a baculovirus expressing recombinant CD1d protein complexed with alpha-galactosylceramide (alpha-GalCer) and quantitated hCD1d tetramer reactive cells in blood and liver from controls and patients with primary biliary cirrhosis (PBC). RESULTS: The majority of CD1d-alphaGalCer-restricted NKT cells were positive for TCR Valpha24 and Vbeta11. There was a distinct CD4- CD8+ population within the CD1d-alphaGalCer-restricted NKT cells in addition to the CD4- CD8- and CD4+ CD8- population. The frequency of CD1d-alphaGalCer-restricted NKT cells was similar between blood and liver in healthy individuals. In contrast, the frequency of CD1d-alphaGalCer-restricted NKT cells in the liver was significantly higher than in the blood of PBC patients. The frequency of CD1d-alpha-GalCer-restricted NKT cells in the liver was also significantly higher in PBC patients than in healthy individuals. CONCLUSIONS: The frequency and function of such cells should be studied not only in blood but also in the target organ of the autoimmune disease. Selective enrichment of CD1d-alphaGalCer-restricted NKT cells at the site of inflammation is observed in PBC, suggesting a role of these cells in the development of PBC. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12360465 ID - 74 ER - TY - JOUR AU - Kita, H. AU - Matsumura, S. AU - He, X. S. AU - Ansari, A. A. AU - Lian, Z. X. AU - Van de Water, J. AU - Coppel, R. L. AU - Kaplan, M. M. AU - Gershwin, M. E. PY - 2002 TI - Quantitative and functional analysis of PDC-E2-specific autoreactive cytotoxic T lymphocytes in primary biliary cirrhosis SP - 1231-40 N1 - May JF - J Clin Invest JO - The Journal of clinical investigation VL - 109 IS - 9 SN - 0021-9738 (Print) N1 - Quantitative and functional analysis of PDC-E2-specific autoreactive cytotoxic T lymphocytes in primary biliary cirrhosis N1 - 11994412 N1 - Dk-39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Autoantigens/*immunology Cells, Cultured Dihydrolipoyllysine-Residue Acetyltransferase Epitopes/immunology Flow Cytometry HLA-A Antigens/immunology Humans Interferon Type II/biosynthesis Liver/immunology Liver Cirrhosis, Biliary/*immunology Pyruvate Dehydrogenase Complex/*immunology T-Lymphocytes, Cytotoxic/*immunology N2 - While the pathologic mechanisms responsible for organ-specific tissue damage in primary biliary cirrhosis (PBC) remain an enigma, it has been suggested that the pathology is mediated by autoreactive T cells infiltrating the intrahepatic bile ducts. Previously, we have documented that there is 100-fold enrichment in the frequency of CD4(+) autoreactive T cells in the liver that are specific for peptides encoded by the E2 components of the pyruvate dehydrogenase complexes (PDC-E2). We have also recently characterized the first MHC class I-restricted epitope for PDC-E2, namely amino acid 159-167, a region very similar to the epitope recognized by MHC class II-restricted CD4(+) cells and by autoantibodies. The effector functions of these PDC-E2(159-167)-specific CD8(+) cytotoxic T lymphocytes (CTLs) are not well understood. We have taken advantage of tetramer technology and report herein that there is tenfold increase in the frequency of PDC-E2(159-167)-specific CTLs in the liver as compared with the blood in PBC. In addition, the precursor frequency of the CTLs in blood was significantly higher in early-stage PBC. Of interest was the fact that, upon stimulation with the peptide, the response of PDC-E2(159-167) tetramer-positive cells is heterogeneous with respect to IFN-gamma synthesis. These data, we believe for the first time, document the enrichment of autoantigen-specific CD8(+) T cells in the PBC liver, suggesting that CD8(+) T cells play a significant role in the immunopathogenesis of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Davis, California 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11994412 ID - 81 ER - TY - JOUR AU - Kita, H. AU - Matsumura, S. AU - He, X. S. AU - Ansari, A. A. AU - Lian, Z. X. AU - Van de Water, J. AU - Coppel, R. L. AU - Kaplan, M. M. AU - Gershwin, M. E. PY - 2002 TI - Analysis of TCR antagonism and molecular mimicry of an HLA-A0201-restricted CTL epitope in primary biliary cirrhosis SP - 918-26 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 36 IS - 4 Pt 1 SN - 0270-9139 (Print) N1 - Analysis of TCR antagonism and molecular mimicry of an HLA-A0201-restricted CTL epitope in primary biliary cirrhosis N1 - 12297839 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Bacterial Proteins/immunology/metabolism CD8-Positive T-Lymphocytes/chemistry/cytology/immunology Cell Line Dihydrolipoyllysine-Residue Acetyltransferase Epitopes Epitopes, T-Lymphocyte/immunology/metabolism Flow Cytometry HLA-A Antigens/*chemistry/immunology/metabolism Humans Interferon Type II/biosynthesis Liver Cirrhosis, Biliary/*immunology Molecular Mimicry/*immunology Peptide Fragments/metabolism Protein Binding/immunology Pseudomonas aeruginosa/chemistry Pyruvate Dehydrogenase Complex/chemistry/immunology/metabolism Receptors, Antigen, T-Cell/antagonists & inhibitors/*chemistry/metabolism T-Lymphocytes, Cytotoxic/*chemistry/cytology/immunology N2 - Although the etiology and mechanism of primary biliary cirrhosis (PBC) is unknown, growing evidence suggests a major role for T cells. We have recently identified the first CD8 T-cell epitope, amino acid 159-167 of the E2 component of pyruvate dehydrogenase complexes (PDC-E2). To seek for analogue peptide-antagonizing effector function of CTLs specific for this autoantigen, we examined the effector functions of the PDC-E2-specific CTLs against alanine substituted peptides. Furthermore, because molecular mimicry has been postulated as a possible cause of initiating PBC, we carried out studies aimed at identifying naturally occurring peptides for the 159-167 peptide of PDC-E2 that may serve as agonists. An alanine substitution at position 5 of this epitope significantly reduced peptide-specific effector functions of CTLs. Moreover, this analogue peptide inhibited effector functions of the CTLs to the prototype peptide, including cytotoxicity and IFN-gamma production. We also identified a peptide derived from Pseudomonas aeruginosa, which showed a higher binding affinity to the HLA-A*0201 than the prototype peptide. This homologous peptide was recognized by CTLs specific for the prototype epitope on PDC-E2. In conclusion, a modification of the immunodominant autoepitope can be utilized to manipulate the CD8 T-cell responses against the autoantigen PDC-E2. Our finding also supports the thesis that molecular mimicry may be implicated in the initiation of the autoreactive CD8 T-cell responses and has implications for the use of such peptides for immunotherapy. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12297839 ID - 75 ER - TY - JOUR AU - Kita, H. AU - Lian, Z. X. AU - Van de Water, J. AU - He, X. S. AU - Matsumura, S. AU - Kaplan, M. AU - Luketic, V. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2002 TI - Identification of HLA-A2-restricted CD8(+) cytotoxic T cell responses in primary biliary cirrhosis: T cell activation is augmented by immune complexes cross-presented by dendritic cells SP - 113-23 N1 - Jan 7 JF - J Exp Med JO - The Journal of experimental medicine VL - 195 IS - 1 SN - 0022-1007 (Print) N1 - Identification of HLA-A2-restricted CD8(+) cytotoxic T cell responses in primary biliary cirrhosis: T cell activation is augmented by immune complexes cross-presented by dendritic cells N1 - 11781370 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - *Antigen Presentation Antigen-Antibody Complex/immunology Autoimmunity Cytokines/secretion Dendritic Cells/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Epitopes HLA-A2 Antigen/*immunology Humans Lymphocyte Activation Oligopeptides/immunology Pyruvate Dehydrogenase Complex/*immunology T-Lymphocytes, Cytotoxic/*immunology N2 - Primary biliary cirrhosis (PBC) is characterized by an intense biliary inflammatory CD4(+) and CD8(+) T cell response. Very limited information on autoantigen-specific cytotoxic T lymphocyte (CTL) responses is available compared with autoreactive CD4(+) T cell responses. Using peripheral blood mononuclear cells (PBMCs) from PBC, we identified an HLA-A2-restricted CTL epitope of the E2 component of pyruvate dehydrogenase (PDC-E2), the immunodominant mitochondrial autoantigen. This peptide, amino acids 159-167 of PDC-E2, induces specific MHC class I-restricted CD8(+) CTL lines from 10/12 HLA-A2(+) PBC patients, but not controls, after in vitro stimulation with antigen-pulsed dendritic cells (DCs). PDC-E2-specific CTLs could also be generated by pulsing DCs with full-length recombinant PDC-E2 protein. Furthermore, using soluble PDC-E2 complexed with either PDC-E2-specific human monoclonal antibody or affinity-purified autoantibodies against PDC-E2, the generation of PDC-E2-specific CTLs, occurred at 100-fold and 10-fold less concentration, respectively, compared with soluble antigen alone. Collectively, these data demonstrate that autoantibody, helper, and CTL epitopes all contain a shared peptide sequence. The finding that autoantigen-immune complexes can not only cross-present but also that presentation of the autoantigen is of a higher relative efficiency, for the first time defines a unique role for autoantibodies in the pathogenesis of an autoimmune disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11781370 ID - 88 ER - TY - JOUR AU - Kimura, Y. AU - Leung, P. S. AU - Kenny, T. P. AU - Van De Water, J. AU - Nishioka, M. AU - Giraud, A. S. AU - Neuberger, J. AU - Benson, G. AU - Kaul, R. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2002 TI - Differential expression of intestinal trefoil factor in biliary epithelial cells of primary biliary cirrhosis SP - 1227-35 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 36 IS - 5 SN - 0270-9139 (Print) N1 - Differential expression of intestinal trefoil factor in biliary epithelial cells of primary biliary cirrhosis N1 - 12395334 N1 - Journal Article United States N1 - eng KW - Biliary Tract/*pathology Biopsy Epithelial Cells/chemistry/pathology/*physiology Gene Expression Regulation Growth Substances/analysis/*genetics Humans Immunohistochemistry In Situ Hybridization Liver Cirrhosis, Biliary/pathology/*physiopathology *Mucins *Muscle Proteins *Neuropeptides Peptides/analysis/*genetics Polymerase Chain Reaction/methods RNA, Messenger/analysis N2 - Intestinal trefoil factor (ITF) promotes epithelial cell migration and mucosal restitution during inflammation. We used real-time quantitative PCR, in situ nucleic acid hybridization, and immunohistochemistry to study the expression of the ITF gene and protein expression in the liver of primary biliary cirrhosis (PBC) and controls. There were significantly higher levels of ITF messenger RNA (mRNA) in PBC liver compared with primary sclerosing cholangitis (PSC) (P <.05) or normal controls (P <.001) and also higher in hepatitis C virus (HCV) liver (P <.05) and cryptogenic cirrhosis (P <.01) compared with normal controls. However, only in PBC was there a significant difference between small (interlobular and bile ductules) and large (intrahepatic and septal) bile ducts. Using in situ hybridization, the highest levels of ITF gene expression were localized to the large bile ducts in PBC. This differential expression of ITF was also noted at the protein level. Thus, in PBC, although 92% of large bile ducts expressed the ITF protein, only 2% of small bile ducts (P <.0001) expressed ITF. In contrast, in control livers, 34% of large bile ducts and 13% of small bile ducts expressed ITF. ITF protein is absent in small bile ducts in all stages of PBC. In conclusion, the expression of ITF may play an important role in bile duct damage. In small bile ducts, ITF production in response to damage is absent, making such cells vulnerable to damage and providing a thesis for the selective loss of small, but not large, bile ducts in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12395334 ID - 71 ER - TY - JOUR AU - Kedzierski, L. AU - Escalante, A. A. AU - Isea, R. AU - Black, C. G. AU - Barnwell, J. W. AU - Coppel, R. L. PY - 2002 TI - Phylogenetic analysis of the genus Plasmodium based on the gene encoding adenylosuccinate lyase SP - 297-301 N1 - Jul JF - Infect Genet Evol VL - 1 IS - 4 SN - 1567-1348 (Print) N1 - Phylogenetic analysis of the genus Plasmodium based on the gene encoding adenylosuccinate lyase N1 - 12798008 N1 - R01 gm60740-01/gm/nigms Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands journal of molecular epidemiology and evolutionary genetics in infectious diseases N1 - eng KW - Adenylosuccinate Lyase/chemistry/*genetics Amino Acid Sequence Animals Evolution, Molecular *Genes, Protozoan Humans Imaging, Three-Dimensional Models, Molecular Molecular Sequence Data *Phylogeny Plasmodium/*genetics/isolation & purification/pathogenicity Poisson Distribution Protein Conformation Sequence Analysis, DNA Sequence Homology, Amino Acid Variation (Genetics) N2 - Phylogenetic studies of the genus Plasmodium have been performed using sequences of the nuclear, mitochondrial and plastid genes. Here we have analyzed the adenylosuccinate lyase (ASL) gene, which encodes an enzyme involved in the salvage of host purines needed by malaria parasites for DNA synthesis. The ASL gene is present in several eukaryotic as well as prokaryotic organisms and does not have repeat regions, which facilitates the accuracy of the alignment. Furthermore, it has been shown that ASL is not subject to positive natural selection. We have sequenced the ASL gene of several different Plasmodium species infecting humans, rodents, monkeys and birds and used the obtained sequences along with the previously known P. falciparum ASL sequence, for structural and phylogenetic analysis of the genus Plasmodium. The genetic divergence of ASL is comparable with that observed in other nuclear genes such as cysteine proteinase, although ASL cannot be considered conserved when compared to aldolase or superoxide dismutase, which exhibit a slower rate of evolution. Nevertheless, a protein like ASL has a rate of evolution that provides enough information for elucidating evolutionary relationships. We modeled 3D structures of the ASL protein based on sequences used in the phylogenetic analysis and obtained a consistent structure for four different species despite the divergence observed. Such models would facilitate alignment in further studies with a greater number of plasmodial species or other Apicomplexa. AD - Department of Microbiology, PO Box 53, Monash University, 3800 Vic, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12798008 ID - 62 ER - TY - JOUR AU - Kedzierski, L. AU - Black, C. G. AU - Goschnick, M. W. AU - Stowers, A. W. AU - Coppel, R. L. PY - 2002 TI - Immunization with a combination of merozoite surface proteins 4/5 and 1 enhances protection against lethal challenge with Plasmodium yoelii SP - 6606-13 N1 - Dec JF - Infect Immun JO - Infection and immunity VL - 70 IS - 12 SN - 0019-9567 (Print) N1 - Immunization with a combination of merozoite surface proteins 4/5 and 1 enhances protection against lethal challenge with Plasmodium yoelii N1 - 12438332 N1 - Journal Article Research Support, Non-U.S. Gov't United States N1 - eng KW - Animals Antibodies, Protozoan/blood Antigens, Protozoan/genetics/*immunology Female Immunization Malaria/parasitology/*prevention & control Membrane Proteins/genetics/immunology Merozoite Surface Protein 1/genetics/immunology Mice Mice, Inbred BALB C Mice, Inbred C3H Plasmodium yoelii/*immunology Protozoan Proteins/genetics/immunology Protozoan Vaccines/*administration & dosage/*immunology Recombinant Proteins/genetics/immunology N2 - It is widely believed that subunit vaccines composed of multiple components will offer greater protection against challenge by malaria, and yet there is little experimental evidence to support this view. We set out to test this proposition in the Plasmodium yoelii challenge system in rodents by comparing the degree of protection conferred by immunization with a mixture of merozoite surface proteins to that conferred by single proteins. We therefore examined a defined protein mixture made of the epidermal growth factor-like domains of P. yoelli merozoite surface protein 1 (MSP1) and MSP4/5, the homologue of P. falciparum MSP4 and MSP5. In the present study we demonstrate that this combination of recombinant proteins dramatically enhances protection against lethal malaria challenge compared to either protein administered alone. Many mice immunized with the MSP4/5 plus MSP1(19) combination did not develop detectable parasitemia after challenge. Combined immunization with MSP1(19) and yMSP4/5, a product characterized by lower protective efficacy, also greatly enhanced protection by reducing peak parasitemias and increasing the numbers of survivors. In some combination trials, levels of antibodies to MSP1(19) were elevated compared to the MSP1(19) alone group; however, improved protection occurred regardless of whether boosting of the anti-MSP1(19) response was observed. Boosting of anti-MSP1(19) did not appear to be due to contaminating endotoxin in the EcMSP4/5 material since enhanced protection was observed in C3H/HeJ mice, which are endotoxin insensitive. Collectively, these experiments show that multiantigen combinations offer enhanced levels of protection against asexual stage infection and suggest that combinations of MSP1, MSP4, and MSP5 should be evaluated further for use in humans. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12438332 ID - 70 ER - TY - JOUR AU - Glenister, F. K. AU - Coppel, R. L. AU - Cowman, A. F. AU - Mohandas, N. AU - Cooke, B. M. PY - 2002 TI - Contribution of parasite proteins to altered mechanical properties of malaria-infected red blood cells SP - 1060-3 N1 - Feb 1 JF - Blood JO - Blood VL - 99 IS - 3 SN - 0006-4971 (Print) N1 - Contribution of parasite proteins to altered mechanical properties of malaria-infected red blood cells N1 - 11807013 N1 - Dk 32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Animals Biomechanics Elasticity/drug effects Erythrocyte Membrane/drug effects/parasitology Erythrocytes/*parasitology/pathology Humans Malaria, Falciparum/*blood Membrane Proteins/pharmacology Peptides/pharmacology Plasmodium falciparum/chemistry/genetics Protozoan Proteins/genetics/*pharmacology N2 - Red blood cells (RBCs) parasitized by Plasmodium falciparum are rigid and poorly deformable and show abnormal circulatory behavior. During parasite development, knob-associated histidine-rich protein (KAHRP) and P falciparum erythrocyte membrane protein 3 (PfEMP3) are exported from the parasite and interact with the RBC membrane skeleton. Using micropipette aspiration, the membrane shear elastic modulus of RBCs infected with transgenic parasites (with kahrp or pfemp3 genes deleted) was measured to determine the contribution of these proteins to the increased rigidity of parasitized RBCs (PRBCs). In the absence of either protein, the level of membrane rigidification was significantly less than that caused by the normal parental parasite clone. KAHRP had a significantly greater effect on rigidification than PfEMP3, contributing approximately 51% of the overall increase that occurs in PRBCs compared to 15% for PfEMP3. This study provides the first quantitative information on the contribution of specific parasite proteins to altered mechanical properties of PRBCs. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11807013 ID - 86 ER - TY - JOUR AU - Fukushima, N. AU - Nalbandian, G. AU - Van De Water, J. AU - White, K. AU - Ansari, A. A. AU - Leung, P. AU - Kenny, T. AU - Kamita, S. G. AU - Hammock, B. D. AU - Coppel, R. L. AU - Stevenson, F. AU - Ishibashi, H. AU - Gershwin, M. E. PY - 2002 TI - Characterization of recombinant monoclonal IgA anti-PDC-E2 autoantibodies derived from patients with PBC SP - 1383-92 N1 - Dec JF - Hepatology JO - Hepatology (Baltimore, Md VL - 36 IS - 6 SN - 0270-9139 (Print) N1 - Characterization of recombinant monoclonal IgA anti-PDC-E2 autoantibodies derived from patients with PBC N1 - 12447863 N1 - Journal Article United States N1 - eng KW - Animals Antibodies, Monoclonal/genetics/*immunology Antibody Specificity Autoantibodies/immunology Baculoviridae/genetics Cell Line Cloning, Molecular Dihydrolipoyllysine-Residue Acetyltransferase Epithelial Cells/cytology/immunology Humans Immunoglobulin A/genetics/*immunology Immunohistochemistry Liver Cirrhosis, Biliary/*immunology Pyruvate Dehydrogenase Complex/*immunology Recombinant Proteins/genetics/immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the presence of autoantibodies to mitochondria (AMA). Recent evidence suggests that PBC develops after a locally driven response in the mucosa, where immunoglobulin A (IgA) is the dominant antibody isotype. In this study, we produced recombinant pyruvate dehydrogenase complex (PDC-E2)-specific dimeric human IgA monoclonal antibodies (mAbs) in a baculovirus expression system. By using 2 anti-PDC-E2 IgG mAbs derived from patients with PBC, we constructed 2 recombinant baculoviruses, each containing heavy chains with the Calpha constant region. These were simultaneously co-infected into Sf9 insect cells with recombinant baculovirus containing the J chain. A sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) immunoblotting profile of the IgA using a 6% nonreducing gel verified the dimeric nature of the autoantibodies. Both recombinants retained their original specificity for PDC-E2. In addition, the antibody showed a mitochondrial staining pattern in HEp2 cells and apically stained the biliary epithelial cells (BECs) in the liver of a patient with PBC but not a normal patient. Transcytosis experiments performed using human polymeric immunoglobulin receptor (pIgR) expressing Madine-Darby canine kidney (MDCK) cells showed that one of the recombinants showed a high degree of colocalization with PDC-E2. In conclusion, these data provide further support of the hypothesis that PDC-E2-specific IgA may enter biliary epithelial cells of PBC patients via the pIgR and complex with PDC-E2, thereby potentially contributing to the pathology of BECs. Moreover, this recombinant PDC-E2-specific mAb provides a tool for further determination of the role of anti-PDC-E2 IgA in the pathogenesis of PBC. AD - Division of Rheumatology/Allergy and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12447863 ID - 69 ER - TY - JOUR AU - Eisen, D. P. AU - Saul, A. AU - Fryauff, D. J. AU - Reeder, J. C. AU - Coppel, R. L. PY - 2002 TI - Alterations in Plasmodium falciparum genotypes during sequential infections suggest the presence of strain specific immunity SP - 8-16 N1 - Jul JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 67 IS - 1 SN - 0002-9637 (Print) N1 - Alterations in Plasmodium falciparum genotypes during sequential infections suggest the presence of strain specific immunity N1 - 12363069 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United States N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/chemistry/genetics Gene Amplification Genotype Haplotypes Membrane Proteins/chemistry/genetics Merozoite Surface Protein 1/chemistry/genetics Molecular Sequence Data Plasmodium falciparum/*genetics/immunology Polymerase Chain Reaction Protozoan Proteins/chemistry/genetics Sequence Homology, Amino Acid N2 - Many of the asexual stage Plasmodium falciparum proteins that are the targets of host protective responses are markedly polymorphic. The full repertoire of diversity is not defined for any antigen. Most studies have focused on the genes encoding merozoite surface proteins 1 and 2 (MSP1, MSP2). We explored the extent of diversity of some of the less studied merozoite surface antigens and analyzed the degree of complexity of malaria field isolates by deriving nucleotide sequences of several antigens. We have determined the genotype of apical membrane antigen 1 (AMA1) in a group of 30 field samples, collected over 29 months, from individuals living in an area of intense malaria transmission in Irian Jaya, identifying 14 different alleles. AMA1 genotyping was combined with previously determined MSP2 typing. AMA1 had the greatest power in distinguishing between isolates but methodological problems, especially when mixed infections are present, suggest it is not an ideal typing target. MSP1, MSP3, and glutamate-rich protein genotypes were also determined from a smaller group of samples, and all results were combined to derive an extended antigenic haplotype. Within this subset of 10 patients, nine different genotypes could be discerned; however, five patients were all infected with the same strain. This strain was present in individuals from two separate villages and was still present 12 months later. This strain was predominant at the first time point but had disappeared at the fourth time point. This significant change in malaria genotypes could be due to strain-specific immunity developing in this population. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12363069 ID - 73 ER - TY - JOUR AU - Cooke, B. M. AU - Glenister, F. K. AU - Mohandas, N. AU - Coppel, R. L. PY - 2002 TI - Assignment of functional roles to parasite proteins in malaria-infected red blood cells by competitive flow-based adhesion assay SP - 203-11 N1 - Apr JF - Br J Haematol JO - British journal of haematology VL - 117 IS - 1 SN - 0007-1048 (Print) N1 - Assignment of functional roles to parasite proteins in malaria-infected red blood cells by competitive flow-based adhesion assay N1 - 11918556 N1 - Dk32094-10/dk/niddk Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Antigens, CD36/metabolism Cell Adhesion Cell Line *Endothelium, Vascular Erythrocyte Membrane/*parasitology Flow Cytometry Gene Deletion Humans Malaria, Falciparum/*blood Membrane Proteins/genetics/metabolism Peptides/genetics/metabolism Plasmodium falciparum/genetics/*metabolism Protozoan Proteins/genetics/*metabolism N2 - Adhesion of parasitized red blood cells (PRBCs) to endothelial cells and subsequent accumulation in the microvasculature are pivotal events in the pathogenesis of falciparum malaria. During intraerythrocytic development, numerous proteins exported from the parasite associate with the RBC membrane skeleton but the precise function of many of these proteins remain unknown. Their cellular location, however, suggests that some may play a role in adhesion. The adhesive properties of PRBCs are best studied under flow conditions in vitro; however, experimental variation in levels of cytoadherence in currently available assays make subtle alterations in adhesion difficult to quantify. Here, we describe a flow-based assay that can quantify small differences in adhesion and document the extent to which a number of parasite proteins influence adhesion using parasite lines that no longer express specific proteins. Loss of parasite proteins ring-infected erythrocyte surface antigen (RESA), knob-associated histidine-rich protein (KAHRP) or Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) had a significant effect on the ability of PRBCs to adhere, whereas loss of mature parasite-infected erythrocyte surface antigen (MESA) had no effect. Our studies indicate that a number of membrane skeleton-associated parasite proteins, although not exposed on the RBC surface, can collectively affect the adhesive properties of PRBCs and further our understanding of pathophysiologically relevant structure/function relationships in malaria-infected RBCs. AD - Department of Microbiology, Monash University, Victoria, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11918556 ID - 83 ER - TY - JOUR AU - Cooke, B. M. AU - Coppel, R. L. AU - Nash, G. B. PY - 2002 TI - Analysis of the adhesive properties of Plasmodium falciparum-infected red blood cells under conditions of flow SP - 561-9 JF - Methods Mol Med JO - Methods in molecular medicine VL - 72 SN - 1543-1894 (Print) N1 - Analysis of the adhesive properties of Plasmodium falciparum-infected red blood cells under conditions of flow N1 - 12125154 N1 - Journal Article United States N1 - eng KW - Animals Antigens, CD/physiology Cell Adhesion/physiology Erythrocytes/*parasitology Host-Parasite Relations Humans Parasitology/methods Plasmodium falciparum/isolation & purification/pathogenicity/*physiology Receptors, IgG/physiology Stress, Mechanical AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12125154 ID - 80 ER - TY - JOUR AU - Cooke, B. M. AU - Coppel, R. L. AU - Nash, G. B. PY - 2002 TI - Preparation of adhesive targets for flow-based cytoadhesion assays SP - 571-9 JF - Methods Mol Med JO - Methods in molecular medicine VL - 72 SN - 1543-1894 (Print) N1 - Preparation of adhesive targets for flow-based cytoadhesion assays N1 - 12125156 N1 - Journal Article United States N1 - eng KW - Antigens, CD/analysis Antigens, CD36/analysis Blood Platelets/cytology/physiology Cell Adhesion/*physiology Cell Culture Techniques/methods Cell Separation/methods Endothelium, Vascular/cytology/*physiology Flow Cytometry/methods Humans Melanoma Tumor Cells, Cultured Umbilical Veins AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12125156 ID - 79 ER - TY - JOUR AU - Black, C. G. AU - Barnwell, J. W. AU - Huber, C. S. AU - Galinski, M. R. AU - Coppel, R. L. PY - 2002 TI - The Plasmodium vivax homologues of merozoite surface proteins 4 and 5 from Plasmodium falciparum are expressed at different locations in the merozoite SP - 215-24 N1 - Apr 9 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 120 IS - 2 SN - 0166-6851 (Print) N1 - The Plasmodium vivax homologues of merozoite surface proteins 4 and 5 from Plasmodium falciparum are expressed at different locations in the merozoite N1 - 11897127 N1 - R01 ai 24710-15/ai/niaid Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Adenylosuccinate Lyase/genetics Amino Acid Sequence Animals Antigens, Protozoan/chemistry/genetics/*metabolism Erythrocytes/parasitology Gene Order/genetics Glycosylphosphatidylinositols Humans Macaca mulatta/parasitology Membrane Proteins/chemistry/genetics/*metabolism Mice Microscopy, Fluorescence Molecular Sequence Data Plasmodium falciparum/genetics/*metabolism/physiology Plasmodium vivax/genetics/*metabolism/physiology Protein Transport Protozoan Proteins/chemistry/genetics/*metabolism Saimiri/parasitology Sequence Homology, Amino Acid Species Specificity N2 - Merozoite surface proteins of Plasmodium falciparum are one major group of antigens currently being investigated and tested as malaria vaccine candidates. Two recently described P. falciparum merozoite surface antigens, MSP4 and MSP5, are GPI-anchored proteins that each contain a single EGF-like domain and appear to have arisen by an ancient gene duplication event. The genes are found in tandem on chromosome 2 of P. falciparum and the syntenic region of the genome was identified in the rodent malarias P. chabaudi, P. yoelii and P. berghei. In these species, there is only a single gene, designated MSP4/5 encoding a single EGF-like domain similar to the EGF-like domain in both PfMSP4 and PfMSP5. Immunization of mice with PyMSP4/5 provides mice with high levels of protection against lethal challenge with blood stage P. yoelii. In this study, we show that in P. vivax, which is quite phylogenetically distant from P. falciparum, both MSP4 and MSP5 homologues can be found with their relative arrangements with respect to the surrounding genes mostly preserved. However, the gene for MSP2, found between MSP5 and adenylosuccinate lyase (ASL) in P. falciparum, is absent from P. vivax. The PvMSP4 and PvMSP5 genes have a two-exon structure and encode proteins with potential signal and GPI anchor sequences and a single EGF-like domain near the carboxyl-terminus. Rabbit antisera raised against purified recombinant proteins show that each of the antisera react with distinct proteins of 62 kDa for PvMSP4 and 86 kDa for PvMSP5 in parasite lysates. Indirect immunofluorescence assays (IFA) localized PvMSP4 over the entire surface of P. vivax merozoites, as expected, whereas, the MSP5 homologue was found to be associated with an apical organellar location consistent with micronemes or over the polar prominence. AD - Department of Microbiology, Monash University, PO Box 53, Calyton 3800 Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11897127 ID - 85 ER - TY - JOUR AU - Bahl, A. AU - Brunk, B. AU - Coppel, R. L. AU - Crabtree, J. AU - Diskin, S. J. AU - Fraunholz, M. J. AU - Grant, G. R. AU - Gupta, D. AU - Huestis, R. L. AU - Kissinger, J. C. AU - Labo, P. AU - Li, L. AU - McWeeney, S. K. AU - Milgram, A. J. AU - Roos, D. S. AU - Schug, J. AU - Stoeckert, C. J., Jr. PY - 2002 TI - PlasmoDB: the Plasmodium genome resource. An integrated database providing tools for accessing, analyzing and mapping expression and sequence data (both finished and unfinished) SP - 87-90 N1 - Jan 1 JF - Nucleic Acids Res JO - Nucleic acids research VL - 30 IS - 1 SN - 1362-4962 (Electronic) N1 - PlasmoDB: the Plasmodium genome resource. An integrated database providing tools for accessing, analyzing and mapping expression and sequence data (both finished and unfinished) N1 - 11752262 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Chromosome Mapping DNA, Protozoan/genetics Database Management Systems *Databases, Genetic Forecasting Gene Expression Profiling *Genome, Protozoan Information Storage and Retrieval Internet Plasmodium/*genetics/*metabolism Plasmodium falciparum/genetics Protozoan Proteins/biosynthesis/genetics Sequence Analysis Sequence Homology N2 - PlasmoDB (http://PlasmoDB.org) is the official database of the Plasmodium falciparum genome sequencing consortium. This resource incorporates finished and draft genome sequence data and annotation emerging from Plasmodium sequencing projects. PlasmoDB currently houses information from five parasite species and provides tools for cross-species comparisons. Sequence information is also integrated with other genomic-scale data emerging from the Plasmodium research community, including gene expression analysis from EST, SAGE and microarray projects. The relational schemas used to build PlasmoDB [Genomics Unified Schema (GUS) and RNA Abundance Database (RAD)] employ a highly structured format to accommodate the diverse data types generated by sequence and expression projects. A variety of tools allow researchers to formulate complex, biologically based queries of the database. A version of the database is also available on CD-ROM (Plasmodium GenePlot), facilitating access to the data in situations where Internet access is difficult (e.g. by malaria researchers working in the field). The goal of PlasmoDB is to enhance utilization of the vast quantities of data emerging from genome-scale projects by the global malaria research community. AD - Department of Biology, University of Pennsylvania, 415 South University Avenue, Philadelphia, PA 19104-6018, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11752262 ID - 91 ER - TY - JOUR AU - Weisman, S. AU - Wang, L. AU - Billman-Jacobe, H. AU - Nhan, D. H. AU - Richie, T. L. AU - Coppel, R. L. PY - 2001 TI - Antibody responses to infections with strains of Plasmodium falciparum expressing diverse forms of merozoite surface protein 2 SP - 959-67 N1 - Feb JF - Infect Immun JO - Infection and immunity VL - 69 IS - 2 SN - 0019-9567 (Print) N1 - Antibody responses to infections with strains of Plasmodium falciparum expressing diverse forms of merozoite surface protein 2 N1 - 11159991 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United States N1 - eng KW - Adolescent Adult Amino Acid Sequence Animals Antibodies, Protozoan/*blood Antigens, Protozoan/chemistry/*immunology Child Enzyme-Linked Immunosorbent Assay Female Humans Malaria, Falciparum/*immunology Male Middle Aged Molecular Sequence Data Plasmodium falciparum/*immunology Protozoan Proteins/chemistry/*immunology N2 - Individuals living in areas where Plasmodium falciparum is endemic experience numerous episodes of infection. These episodes may or may not be symptomatic, with the outcome depending on a combination of parasite and host factors, several of which are poorly understood. One factor is believed to be the particular alleles of several parasite proteins to which the host is capable of mounting protective immune responses. We report a study examining antibody responses to MSP2 in 15 semi-immune teenagers and adults living in the Khanh-Hoa area of southern-central Vietnam, where P. falciparum is highly endemic; subjects were serially infected with multiple strains of P. falciparum. The MSP2 alleles infecting these subjects were determined by nucleotide sequencing. A total of 62 MSP2 genes belonging to both dimorphic families were identified, of which 33 contained distinct alleles, with 61% of the alleles being detected once. Clear changes in the repertoire occurred between infections. Most infections contained a mixture of parasites expressing MSP2 alleles from both dimorphic families. Two examples of reinfection with a strain expressing a previously encountered allele were detected. Significant changes in antibody levels to various regions of MSP2 were detected over the course of the experiment. There was no clear relation between the infecting form of MSP2 and the ensuing antibody response. This study highlights the complexity of host-parasite relationship for this important human pathogen. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11159991 ID - 107 ER - TY - JOUR AU - Wang, L. AU - Richie, T. L. AU - Stowers, A. AU - Nhan, D. H. AU - Coppel, R. L. PY - 2001 TI - Naturally acquired antibody responses to Plasmodium falciparum merozoite surface protein 4 in a population living in an area of endemicity in Vietnam SP - 4390-7 N1 - Jul JF - Infect Immun JO - Infection and immunity VL - 69 IS - 7 SN - 0019-9567 (Print) N1 - Naturally acquired antibody responses to Plasmodium falciparum merozoite surface protein 4 in a population living in an area of endemicity in Vietnam N1 - 11401978 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United States N1 - eng KW - Adolescent Adult Animals Antibodies, Protozoan/*blood/immunology Antibody Specificity Antigens, Protozoan/*immunology Child *Endemic Diseases Epitopes, B-Lymphocyte/immunology Humans Immunoglobulin Isotypes Malaria, Falciparum/blood/epidemiology/*immunology/prevention & control Merozoite Surface Protein 1/immunology Middle Aged Plasmodium falciparum/immunology Prevalence Protozoan Proteins/*immunology Time Factors Vietnam/epidemiology N2 - Merozoite surface protein 4 (MSP4) of Plasmodium falciparum is a glycosylphosphatidylinositol-anchored integral membrane protein that is being developed as a component of a subunit vaccine against malaria. We report here the measurement of naturally acquired antibodies to MSP4 in a population of individuals living in the Khanh-Hoa region of Vietnam, an area where malaria is highly endemic. Antibodies to MSP4 were detected in 94% of the study population at titers of 1:5,000 or greater. Two forms of recombinant MSP4 produced in either Escherichia coli or Saccharomyces cerevisiae were compared as substrates in the enzyme-linked immunosorbent assay. There was an excellent correlation between reactivity measured to either, although the yeast substrate was recognized by a higher percentage of sera. Four different regions of MSP4 were recognized by human antibodies, demonstrating that there are at least four distinct epitopes in this protein. In the carboxyl terminus, where the single epidermal growth factor-like domain is located, the reactive epitope(s) was shown to be conformation dependent, as disruption of the disulfide bonds almost completely abolished reactivity with human antibodies. The anti-MSP4 antibodies were mainly of the immunoglobulin G1 (IgG1) and IgG3 subclasses, suggesting that such antibodies may play a role in opsonization and complement-mediated lysis of free merozoites. Individuals in the study population were drug-cured and followed up for 6 months; no significant correlation was observed between the anti-MSP4 antibodies and the absence of parasitemia during the surveillance period. As a comparison, antibodies to MSP1(19), a leading vaccine candidate, were measured, and no correlation with protection was observed in these individuals. The anti-MSP1(19) antibodies were predominantly of the IgG1 isotype, in contrast to the IgG3 predominance noted for MSP4. AD - Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11401978 ID - 100 ER - TY - JOUR AU - Van de Water, J. AU - Ishibashi, H. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2001 TI - Molecular mimicry and primary biliary cirrhosis: premises not promises SP - 771-5 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 33 IS - 4 SN - 0270-9139 (Print) N1 - Molecular mimicry and primary biliary cirrhosis: premises not promises N1 - 11283838 N1 - Journal Article Review United States N1 - eng KW - Bile Ducts/cytology/immunology Epithelial Cells/immunology Hepatitis, Autoimmune/immunology Humans Liver Cirrhosis, Biliary/genetics/*immunology *Molecular Mimicry T-Lymphocytes/physiology Xenobiotics/immunology AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11283838 ID - 105 ER - TY - JOUR AU - Uhlemann, A. C. AU - Oguariri, R. M. AU - McColl, D. J. AU - Coppel, R. L. AU - Kremsner, P. G. AU - Anders, R. F. AU - Kun, J. F. PY - 2001 TI - Properties of the Plasmodium falciparum homologue of a protective vaccine candidate of Plasmodium yoelii SP - 41-8 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 118 IS - 1 SN - 0166-6851 (Print) N1 - Properties of the Plasmodium falciparum homologue of a protective vaccine candidate of Plasmodium yoelii N1 - 11704272 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Base Sequence Malaria/prevention & control Malaria Vaccines/*genetics/immunology Membrane Proteins/chemistry/*genetics/immunology Molecular Sequence Data Plasmodium falciparum/*immunology Plasmodium yoelii/*immunology Protozoan Proteins/chemistry/*genetics/immunology Sequence Analysis, DNA Sequence Homology, Amino Acid N2 - We describe an unusual tryptophan-rich protein of Plasmodium falciparum that contains threonine-rich repeats. The protein is encoded by a 2.5 kb gene with a two-exon structure including a short AT-rich intron that is spliced out of the mature message. The 5' end of the gene encodes a hydrophobic region, which is assumed to be a signal peptide. The peptide sequence is characterised by a tryptophan-rich region and a block of degenerate threonine repeats. The protein is synthesised throughout the asexual life cycle and has an apparent molecular weight of approximately 94 kDa. It has a variable molecular weight in different strains of P. falciparum. Length polymorphisms can be found in the intron region and the second exon. Four single nucleotide mutations are localised in the tryptophan-rich region and two were found in the threonine-repeat block. Homology searches based on gene structure and amino acid sequence revealed a relationship with a P. yoelii antigen that has been used successfully in vaccine studies. Thus, this P. falciparum antigen should be considered an additional candidate for assessment in vaccination against the asexual blood-stages of P. falciparum. AD - Department of Parasitology, Institute of Tropical Medicine, University of Tubingen, Wilhelmstrasse 27, 72074, Tubingen, Germany. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11704272 ID - 95 ER - TY - JOUR AU - Tanaka, A. AU - Leung, P. S. AU - Kenny, T. P. AU - Au-Young, J. AU - Prindiville, T. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2001 TI - Genomic analysis of differentially expressed genes in liver and biliary epithelial cells of patients with primary biliary cirrhosis SP - 89-98 N1 - Aug JF - J Autoimmun JO - Journal of autoimmunity VL - 17 IS - 1 SN - 0896-8411 (Print) N1 - Genomic analysis of differentially expressed genes in liver and biliary epithelial cells of patients with primary biliary cirrhosis N1 - 11488641 N1 - Dk39588/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Bile Ducts/*metabolism/pathology Epithelial Cells/*metabolism/pathology *Gene Expression Regulation/genetics Gene Library Humans Liver/*metabolism/pathology Liver Cirrhosis, Biliary/*genetics/pathology Nucleic Acid Hybridization/methods RNA, Messenger/biosynthesis Sequence Homology, Nucleic Acid N2 - The characterization of differentially expressed genes provides a powerful tool for identifying molecules that may be involved in the pathogenesis of disease. We have used two independent techniques to identify overexpressed transcripts in bile duct cells and in liver from patients with primary biliary cirrhosis (PBC). In the first method, we used suppressive subtractive hybridization to compare mRNA from isolated PBC bile duct epithelial cells (BECs) to normal BECs and identified 71 clones as transcribed at higher levels in PBC-BECs. Amongst these clones, 62/71 had matches in a non-redundant nucleotide database and 9/71 had matches in an EST database. Of the 62 clones, 51/62 include a complexity of genes involved in cell proliferation, signal transduction, transcription regulation, RNA processing, carbohydrate metabolism and hypothetical/unknown proteins; 4/62 were identified as interstitial collagenase and collagenase precursors, 4/62 as ribosomal proteins, 3/62 as mitochondrial DNA. The mitochondrial cDNA sequences included cytochrome c oxidase, Wnt-13, and the pHL gene, a c-myc oncogene containing coxIII sequence. In the second method, we constructed cDNA libraries from three different PBC livers and sequenced a total of 12,324 independent clones. These 12,324 clones underwent virtual subtraction with 2,814,148 independent clones from Incyte LifeSeq libraries. Twenty one sequences were identified as unique to PBC liver. Collectively, these approaches identified a number of genes involved in signalling, RNA processing, mitochondrial function, inflammation, and fibrosis. Interestingly, both Wnt-13 and Notch transcripts are overexpressed in PBC liver. Further studies are needed to focus on the significance of these genes during the natural history of disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, Department of Internal Medicine, University of California at Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11488641 ID - 99 ER - TY - JOUR AU - Stern, L. AU - Allison, L. AU - Coppel, R. L. AU - Dix, T. I. PY - 2001 TI - Discovering patterns in Plasmodium falciparum genomic DNA SP - 175-86 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 118 IS - 2 SN - 0166-6851 (Print) N1 - Discovering patterns in Plasmodium falciparum genomic DNA N1 - 11738708 N1 - Evaluation Studies Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Algorithms Animals Chromosomes/genetics Computational Biology/*methods DNA, Protozoan/*genetics *Genome, Protozoan *Information Theory Markov Chains Models, Genetic Models, Statistical Plasmodium falciparum/*genetics Probability Repetitive Sequences, Nucleic Acid/*genetics Sequence Analysis, DNA Telomere N2 - A method has been developed for discovering patterns in DNA sequences. Loosely based on the well-known Lempel Ziv model for text compression, the model detects repeated sequences in DNA. The repeats can be forward or inverted, and they need not be exact. The method is particularly useful for detecting distantly related sequences, and for finding patterns in sequences of biased nucleotide composition, where spurious patterns are often observed because the bias leads to coincidental nucleotide matches. We show here the utility of the method by applying it to genomic sequences of Plasmodium falciparum. A single scan of chromosomes 2 and 3 of P. falciparum, using our method and no other a priori information about the sequences, reveals regions of low complexity in both telomeric and central regions, long repeats in the subtelomeric regions, and shorter repeat areas in dense coding regions. Application of the method to a recently sequenced contig of chromosome 10 that has a particularly biased base composition detects a long internal repeat more readily than does the conventional dot matrix plot. Space requirements are linear, so the method can be used on large sequences. The observed repeat patterns may be related to large-scale chromosomal organization and control of gene expression. The method has general application in detecting patterns of potential interest in newly sequenced genomic material. AD - Department of Computer Science and Software Engineering, The University of Melbourne, Melbourne, Victoria 3010, Australia. stern@unimelb.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11738708 ID - 92 ER - TY - JOUR AU - Sasaki, M. AU - Van De Water, J. AU - Kenny, T. P. AU - Gallo, M. L. AU - Leung, P. S. AU - Nakanuma, Y. AU - Ansari, A. A. AU - Coppel, R. L. AU - Neuberger, J. AU - Gershwin, M. E. PY - 2001 TI - Immunoglobulin gene usage and immunohistochemical characteristics of human monoclonal antibodies to the mitochondrial autoantigens of primary biliary cirrhosis induced in the XenoMouse SP - 631-7 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 34 IS - 4 Pt 1 SN - 0270-9139 (Print) N1 - Immunoglobulin gene usage and immunohistochemical characteristics of human monoclonal antibodies to the mitochondrial autoantigens of primary biliary cirrhosis induced in the XenoMouse N1 - 11584357 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence Animals Antibodies, Monoclonal/*immunology Autoantigens/*immunology Dihydrolipoyllysine-Residue Acetyltransferase *Genes, Immunoglobulin Immunization Immunoglobulin Variable Region/genetics Immunohistochemistry Liver Cirrhosis, Biliary/etiology/*immunology Mice Mitochondria/*immunology Molecular Sequence Data Pyruvate Dehydrogenase Complex/*immunology N2 - The immunodominant antimitochondrial antibody (AMA) response in primary biliary cirrhosis (PBC) is directed against the E2 component of pyruvate dehydrogenase (PDC-E2). The nature of the clonal selection process is unclear, and to address this issue, we took advantage of a transgenic technology, XenoMouse, that contains 80% of the human immunoglobulin (Ig) variable gene repertoire and can produce high-affinity human antibodies to virtually any immunogen without evidence of clonal bias. We immunized mice with PDC-E2 to obtain 13 HmAbs, including 4 IgG(2) and 9 IgM isotypes. Immunoglobulin gene analysis was unique and demonstrated a clonal bias; the immunoglobulin gene usage was considerably different from other antibody responses analyzed in XenoMouse systems. Four of the 13 mAbs recognized the inner lipoyl domain of PDC-E2, 2 of 13 recognized the entire PDC-E2 molecule, 4 of 13 recognized PDC-E2 and OGDC-E2, 1 of 13 recognized OGDC only, 1 recognized BCOADC-E2 only, and 1 recognized an unidentified 100-kd mitochondrial protein. Immunohistochemical staining using these HmAbs produced mitochondrial staining of septal bile ducts in both PBC and control livers. Ig gene analysis showed that 7 of 13 HmAbs used the V(H)3 and 4 of 13 used VH4 gene repertoire, respectively. Three of 7 V(H)3 antibodies used the same Ig VH3-21 gene family found in human AMA from patients with PBC. The CDRs of these autoantibodies were slightly mutated when compared with the sequences present within the Ig germline genes. In conclusion, the XenoMouse not only recapitulates the unique specificity and restriction of PBC patients, but indicates that the autoantibodies are derived from a restricted clonal selection process. Such data suggest that the original immunogen leads to somatic mutation without subsequent development of determinant spreading. AD - Division of Rheumatology/Allergy and Clinical Immunology, School of Medicine University of California, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11584357 ID - 96 ER - TY - JOUR AU - Nishio, A. AU - Bass, N. M. AU - Luketic, V. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 2001 TI - Primary biliary cirrhosis: from induction to destruction SP - 89-102 N1 - Apr JF - Semin Gastrointest Dis JO - Seminars in gastrointestinal disease VL - 12 IS - 2 SN - 1049-5118 (Print) N1 - Primary biliary cirrhosis: from induction to destruction N1 - 11352123 N1 - Dk 39588/dk/niddk Es 10319/es/niehs Case Reports Journal Article Research Support, U.S. Gov't, P.H.S. Review United States N1 - eng KW - Adult Female Humans Liver Cirrhosis, Biliary/blood/complications/*diagnosis/immunology Liver Function Tests Liver Transplantation Sjogren's Syndrome/blood/complications/*diagnosis N2 - Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease that predominantly affects middle-aged women; fatigue and pruritus are the most common symptoms at presentation. Liver function tests are consistent with cholestasis and reveal an elevation of serum alkaline phosphatase and gamma-glutamyl transpeptidase with or without elevation of aminotransferase levels. Histologically, PBC is characterized by the destruction of the intrahepatic small bile ducts and subsequently fibrosis. The serological hallmark of the disease is the presence of antimitochondrial antibodies, which are found in 95% of patients with PBC. The antimitochondrial antibodies are directed against the 2-oxo-acid dehydrogenase complexes located on the inner membrane of the mitochondria. PBC generally slowly progresses, even over decades, and may lead to liver failure. In symptomatic patients, advanced age, elevated serum bilirubin levels, decreased serum albumin levels, and cirrhosis each correlate with shortened survival. Immunosuppressive and anti-inflammatory drugs have been used in the treatment of PBC based on the presumed autoimmune pathogenesis, but satisfactory agents leading to complete reversal or cure of the disease are not available. At present ursodeoxycholic acid appears to be the only effective therapy in preventing or delaying the need for liver transplantation and improving survival. However, a number of patients receiving ursodeoxycholic acid still develop progressive disease and require transplantation; transplantation is the only effective therapy at the end stage of the disease. AD - Department of Gastroenterology, Tenri Hospital, Nara, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11352123 ID - 103 ER - TY - JOUR AU - Migliaccio, C. AU - Van de Water, J. AU - Ansari, A. A. AU - Kaplan, M. M. AU - Coppel, R. L. AU - Lam, K. S. AU - Thompson, R. K. AU - Stevenson, F. AU - Gershwin, M. E. PY - 2001 TI - Heterogeneous response of antimitochondrial autoantibodies and bile duct apical staining monoclonal antibodies to pyruvate dehydrogenase complex E2: the molecule versus the mimic SP - 792-801 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 33 IS - 4 SN - 0270-9139 (Print) N1 - Heterogeneous response of antimitochondrial autoantibodies and bile duct apical staining monoclonal antibodies to pyruvate dehydrogenase complex E2: the molecule versus the mimic N1 - 11283841 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United States N1 - eng KW - Amino Acid Sequence/genetics Animals Antibodies, Monoclonal/immunology Autoantibodies/*immunology Bile Ducts/*enzymology Cell Membrane/enzymology Dihydrolipoyllysine-Residue Acetyltransferase Epitopes Mice Mice, Inbred BALB C Mitochondria/*immunology Molecular Mimicry Molecular Sequence Data Pyruvate Dehydrogenase Complex/genetics/*immunology/*metabolism Staining and Labeling N2 - The 2-oxo-acid dehydrogenase complexes and, in particular, the E2 component of the pyruvate dehydrogenase complex (PDC) are the target of antimitochondrial antibodies (AMA). More than 95% of primary biliary cirrhosis (PBC) patients have detectable levels of autoantibodies to PDC-E2 and in general these react with a region of the molecule that contains the prosthetic group lipoic acid (LA). LA is vital to the function of the enzyme, although there is conflicting evidence as to whether its presence is required for PDC-E2 recognition by AMA. Some, but not all, monoclonal antibodies (mAbs) to PDC-E2 produce an intense staining pattern at the apical surface of bile duct epithelial cells (BEC) in patients with PBC, and it has been argued that the molecule at the apical surface of PBC bile duct cells is a modified form of PDC-E2 or a cross-reactive molecule, acting as a molecular mimic. Herein, we characterize the epitopes recognized by 4 anti-PDC-E2 mAbs that give apical staining patterns (3 mouse and 1 human). In particular, by using a combination of recombinant antigens, competitive inhibition assays, and a unique peptide-on-bead assay, we determined that these apically staining mAbs recognize 3 or 4 distinct epitopes on PDC-E2. More importantly, this suggests that a portion spanning the entire inner lipoyl domain of PDC-E2 can be found at the BEC apical surface. In addition, competition assays with patient sera and a PDC-E2-specific mAb showed significant epitope overlap with only 1 of the 3 mouse mAbs and showed a differential response to the peptide bound to beads. These findings further highlight the heterogeneous response of patient autoantibodies to the inner lipoyl domain of PDC-E2. AD - Divisions of Rheumatology, Allergy & Clinical Immunology, and Hematology & Oncology, Department of Internal Medicine, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11283841 ID - 104 ER - TY - JOUR AU - Long, S. A. AU - Quan, C. AU - Van de Water, J. AU - Nantz, M. H. AU - Kurth, M. J. AU - Barsky, D. AU - Colvin, M. E. AU - Lam, K. S. AU - Coppel, R. L. AU - Ansari, A. AU - Gershwin, M. E. PY - 2001 TI - Immunoreactivity of organic mimeotopes of the E2 component of pyruvate dehydrogenase: connecting xenobiotics with primary biliary cirrhosis SP - 2956-63 N1 - Sep 1 JF - J Immunol VL - 167 IS - 5 SN - 0022-1767 (Print) N1 - Immunoreactivity of organic mimeotopes of the E2 component of pyruvate dehydrogenase: connecting xenobiotics with primary biliary cirrhosis N1 - 11509645 N1 - Dk39588/dk/niddk Es103019/es/niehs Journal Article Research Support, U.S. Gov't, P.H.S. United States 1950) N1 - eng KW - Amino Acid Sequence Autoantibodies/biosynthesis Autoantigens/chemistry Epitopes/chemistry Humans Liver Cirrhosis, Biliary/enzymology/*etiology/*immunology *Molecular Mimicry Molecular Structure Peptide Fragments/chemistry/genetics/immunology Pyruvate Dehydrogenase Complex/chemistry/genetics/*immunology Thioctic Acid/chemistry Xenobiotics/chemistry/*immunology/*toxicity N2 - In primary biliary cirrhosis (PBC), the major autoepitope recognized by both T and B cells is the inner lipoyl domain of the E2 component of pyruvate dehydrogenase. To address the hypothesis that PBC is induced by xenobiotic exposure, we took advantage of ab initio quantum chemistry and synthesized the inner lipoyl domain of E2 component of pyruvate dehydrogenase, replacing the lipoic acid moiety with synthetic structures designed to mimic a xenobiotically modified lipoyl hapten, and we quantitated the reactivity of these structures with sera from PBC patients. Interestingly, antimitochondrial Abs from all seropositive patients with PBC, but no controls, reacted against 3 of the 18 organic modified autoepitopes significantly better than to the native domain. By structural analysis, the features that correlated with autoantibody binding included synthetic domain peptides with a halide or methyl halide in the meta or para position containing no strong hydrogen bond accepting groups on the phenyl ring of the lysine substituents, and synthetic domain peptides with a relatively low rotation barrier about the linkage bond. Many chemicals including pharmaceuticals and household detergents have the potential to form such halogenated derivatives as metabolites. These data reflect the first time that an organic compound has been shown to serve as a mimeotope for an autoantigen and further provide evidence for a potential mechanism by which environmental organic compounds may cause PBC. AD - Division of Rheumatology, University of California School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11509645 ID - 98 ER - TY - JOUR AU - Kedzierski, L. AU - Black, C. G. AU - Stowers, A. W. AU - Goschnick, M. W. AU - Kaslow, D. C. AU - Coppel, R. L. PY - 2001 TI - Comparison of the protective efficacy of yeast-derived and Escherichia coli-derived recombinant merozoite surface protein 4/5 against lethal challenge by Plasmodium yoelii SP - 4661-8 N1 - Sep 14 JF - Vaccine JO - Vaccine VL - 19 IS - 32 SN - 0264-410X (Print) N1 - Comparison of the protective efficacy of yeast-derived and Escherichia coli-derived recombinant merozoite surface protein 4/5 against lethal challenge by Plasmodium yoelii N1 - 11535314 N1 - Comparative Study Evaluation Studies Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. England N1 - eng KW - Animals Antibodies, Protozoan/biosynthesis/immunology Antigens, Protozoan/biosynthesis/genetics/*immunology/isolation & purification Escherichia coli/*metabolism Female Immunoglobulin G/biosynthesis/immunology Immunoglobulin Isotypes/biosynthesis/immunology Malaria/immunology/*prevention & control Malaria Vaccines/biosynthesis/genetics/*immunology/isolation & purification Membrane Proteins/biosynthesis/genetics/*immunology/isolation & purification Mice Mice, Inbred BALB C Parasitemia/immunology Plasmodium yoelii/genetics/*immunology Protein Structure, Tertiary Protozoan Proteins/biosynthesis/genetics/*immunology/isolation & purification Recombinant Fusion Proteins/biosynthesis/genetics/immunology/isolation & purification Saccharomyces cerevisiae/*metabolism Species Specificity Structure-Activity Relationship Vaccination N2 - The gene encoding the Plasmodium yoelii homologue of P. falciparum merozoite surface proteins 4 (MSP4) and 5 (MSP5) has been expressed in Escherichia coli and Saccharomyces cerevisiae. The protein contains a single epidermal growth factor (EGF)-like domain and is expressed in a form lacking the predicted N-terminal signal and glycosyl phosphatidylinositol (GPI) attachment sequences. The recombinant protein derived from E. coli (EcMSP4/5) was highly effective at protecting mice against lethal challenge with 10(5) parasites of the P. yoelii YM strain. In contrast, the protective efficacy of yeast-derived MSP4/5 (yMSP4/5) was considerably less. The antibody titres in both groups were significantly different with mice immunised with yeast-derived protein showing significantly lower pre-challenge antibody responses. There was a significant inverse correlation between antibody levels as measured by ELISA and peak parasitaemia. Mice immunised with EcMSP4/5 produced anti-PyMSP4/5 antibodies predominantly of the IgG2a and IgG2b isotypes, whereas, mice immunised with yMSP4/5 mainly produced antibodies of the IgG1 isotype. The differences in antibody titres and subtype distribution may account for the observed differences in protective efficacy of these protein preparations. Levels of protective efficacy of MSP4/5 were compared with that obtained using P. yoelii MSP1 produced in S. cerevisiae. Levels of protection induced by E. coli derived MSP4/5 were superior to those induced by MSP1 which in turn were better than those induced by yeast-derived MSP4/5. AD - Department of Microbiology, Monash University, PO Box 53, Victoria 3800, Clayton, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11535314 ID - 97 ER - TY - JOUR AU - Doolan, D. L. AU - Coppel, R. L. AU - Waters, A. P. PY - 2001 TI - Foreword--MBP thematic issue on genomics SP - 127-8 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 118 IS - 2 SN - 0166-6851 (Print) N1 - Foreword--MBP thematic issue on genomics N1 - 11738701 N1 - Editorial Netherlands N1 - eng KW - Animals Computational Biology/*methods Genome, Protozoan *Genomics Humans Malaria/diagnosis/*prevention & control Malaria Vaccines Plasmodium/*genetics Proteome Protozoan Proteins/genetics/metabolism UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11738701 ID - 94 ER - TY - JOUR AU - Coppel, R. L. PY - 2001 TI - Bioinformatics and the malaria genome: facilitating access and exploitation of sequence information SP - 139-45 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 118 IS - 2 SN - 0166-6851 (Print) N1 - Bioinformatics and the malaria genome: facilitating access and exploitation of sequence information N1 - 11738704 N1 - Journal Article Research Support, Non-U.S. Gov't Review Netherlands N1 - eng KW - Animals *Computational Biology Databases, Genetic *Genome, Protozoan Humans Information Storage and Retrieval Internet Malaria/parasitology Plasmodium falciparum/*genetics N2 - The torrent of sequence information unleashed by the various genome sequencing projects, including that of Plasmodium falciparum, will lead to an unprecedented increase in the data available for research purposes. The scientific community is struggling to develop ways to assimilate this information and ensure that it is fully analysed in a way that enables rapid development of new therapeutic and diagnostic advances. This is particularly so for the field of tropical medicine where many of the scientists have had limited training in the area of Bioinformatics and may be further hampered by poor access to the sequence data. A number of collections of malaria genome sequence are available, each with their own advantages and disadvantages, however further improvements in these information resources are needed. In particular, there would be great benefit in integrating genomic sequence and functional genomics results with the large amount of pre-existing knowledge related to parasite biology and immunological interactions with the host. Attempts to achieve this include the PlasmoDB database, and the lessons learned in this effort could be of great utility to other organism-specific databases. AD - Department of Microbiology and the Victorian Bioinformatics Consortium, P.O. Box 53, Monash University, Melbourne, Victoria 3800, Australia. ross.coppel@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11738704 ID - 93 ER - TY - JOUR AU - Cooke, B. M. AU - Mohandas, N. AU - Coppel, R. L. PY - 2001 TI - The malaria-infected red blood cell: structural and functional changes SP - 1-86 JF - Adv Parasitol JO - Advances in parasitology VL - 50 SN - 0065-308X (Print) N1 - The malaria-infected red blood cell: structural and functional changes N1 - 11757330 N1 - Comparative Study Journal Article Review England N1 - eng KW - Animals Babesiosis/blood/parasitology Biological Transport, Active Blood Proteins/physiology Cell Adhesion Erythrocyte Membrane/parasitology/physiology Erythrocytes/parasitology/*pathology/*physiology Host-Parasite Relations Humans Malaria/*blood/parasitology Plasmodium/genetics/parasitology Protozoan Proteins/blood/genetics Rheology N2 - The asexual stage of malaria parasites of the genus Plasmodium invade red blood cells of various species including humans. After parasite invasion, red blood cells progressively acquire a new set of properties and are converted into more typical, although still simpler, eukaryotic cells by the appearance of new structures in the red blood cell cytoplasm, and new proteins at the red blood cell membrane skeleton. The red blood cell undergoes striking morphological alterations and its rheological properties are considerably altered, manifesting as red blood cells with increased membrane rigidity, reduced deformability and increased adhesiveness for a number of other cells including the vascular endothelium. Elucidation of the structural changes in the red blood cell induced by parasite invasion and maturation and an understanding of the accompanying functional alterations have the ability to considerably extend our knowledge of structure-function relationships in the normal red blood cell. Furthermore, interference with these interactions may lead to previously unsuspected means of reducing parasite virulence and may lead to the development of novel antimalarial therapeutics. AD - Department of Microbiology, P.O. Box 53, Monash University, Victoria 3800, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11757330 ID - 89 ER - TY - JOUR AU - Black, C. G. AU - Wu, T. AU - Wang, L. AU - Hibbs, A. R. AU - Coppel, R. L. PY - 2001 TI - Merozoite surface protein 8 of Plasmodium falciparum contains two epidermal growth factor-like domains SP - 217-26 N1 - May JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 114 IS - 2 SN - 0166-6851 (Print) N1 - Merozoite surface protein 8 of Plasmodium falciparum contains two epidermal growth factor-like domains N1 - 11378201 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan/immunology Antigenic Variation Antigens, Protozoan/*chemistry/genetics Epidermal Growth Factor/chemistry Fluorescent Antibody Technique, Indirect Humans Molecular Sequence Data Plasmodium falciparum/*chemistry/*genetics Polymerase Chain Reaction Protozoan Proteins/*chemistry/genetics Rabbits Recombinant Fusion Proteins/chemistry Sequence Alignment Sequence Homology, Amino Acid N2 - By motif searching of the unfinished sequences in the Malaria Genome Sequencing Project databases we have identified a novel EGF-like domain-containing protein of Plasmodium falciparum. The sequence lies within a single open reading frame of 1791 bp and is predicted to encode a polypeptide of 597 amino acids. There are hydrophobic regions at the extreme N- and C-termini, which could represent secretory signal peptide and GPI attachment sites, respectively. Similar to MSP1, there are two EGF-like domains located near the C-terminus. RT-PCR analysis of the novel gene shows that it is transcribed in asexual stages of the malaria parasite. We have expressed portions of the protein as recombinant GST fusions in Escherichia coli and raised antisera in rabbits. Antibodies to the EGF-like domains of the novel protein are highly specific and do not cross-react with the EGF-like domains of MSP1, MSP4 or MSP5 expressed as GST fusion proteins. Antiserum raised to the most C-terminal region of the protein reacts with four bands of 98, 50, 25 and 19 kDa in P. falciparum parasite lysates whereas antisera to the N-terminal fusion proteins recognise the 98 and 50 kDa bands, suggesting that the novel protein may undergo processing in a similar way to MSP1. Immunoblot analysis of stage-specific parasite samples reveals that the protein is present throughout the parasite asexual life cycle and in isolated merozoites, with the smaller fragments present in ring stage parasites. The protein partitions in the detergent-enriched phase after Triton X-114 fractionation and is localized to the surfaces of trophozoites, schizonts and free merozoites by indirect immunofluorescence. Antisera to the C-terminus stain the surface of rings, whereas antisera to the N-terminus do not, suggesting that a fragment of the protein is carried into the developing ring stage parasite. Based on the accepted nomenclature in the field we designate this protein MSP8. We have shown that the MSP8 fusion proteins are in a conformation that can be recognised by human immune sera and that there is very limited diversity in the MSP8 gene sequences from various P. falciparum laboratory isolates. MSP8 shows significant similarity to the recently reported sequence of the protective P. yoelii merozoite surface protein pypAg-2 [Burns JM, Belk CC, Dunn PD. Infect Immun 2000;68:6189-95.] suggesting that the two proteins are homologues. Taken together, these findings suggest that MSP8/pypAg-2 may play an important role in the process of red cell invasion and is a potential malaria vaccine candidate. AD - Department of Microbiology, PO Box 53, Monash University, 3800, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11378201 ID - 101 ER - TY - JOUR AU - Waterkeyn, J. G. AU - Wickham, M. E. AU - Davern, K. M. AU - Cooke, B. M. AU - Coppel, R. L. AU - Reeder, J. C. AU - Culvenor, J. G. AU - Waller, R. F. AU - Cowman, A. F. PY - 2000 TI - Targeted mutagenesis of Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) disrupts cytoadherence of malaria-infected red blood cells SP - 2813-23 N1 - Jun 15 JF - Embo J JO - The EMBO journal VL - 19 IS - 12 SN - 0261-4189 (Print) N1 - Targeted mutagenesis of Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) disrupts cytoadherence of malaria-infected red blood cells N1 - 10856227 N1 - 1ro1ai44008/ai/niaid Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Antigens, CD36/metabolism Biological Transport Cell Adhesion Cell Compartmentation Cell Polarity Endothelium, Vascular/parasitology Erythrocyte Membrane/*parasitology/ultrastructure Genes, Protozoan Membrane Proteins/*genetics/metabolism Mutagenesis Peptides/metabolism Plasmodium falciparum/*genetics/ultrastructure Protozoan Proteins/metabolism Recombinant Proteins/biosynthesis N2 - Adhesion of parasite-infected red blood cells to the vascular endothelium is a critical event in the pathogenesis of malaria caused by Plasmodium falciparum. Adherence is mediated by the variant erythrocyte membrane protein 1 (PfEMP1). Another protein, erythrocyte membrane protein-3 (PfEMP3), is deposited under the membrane of the parasite-infected erythrocyte but its function is unknown. Here we show that mutation of PfEMP3 disrupts transfer of PfEMP1 to the outside of the P.FALCIPARUM:-infected cell. Truncation of the C-terminal end of PfEMP3 by transfection prevents distribution of this large (>300 kDa) protein around the membrane but does not disrupt trafficking of the protein from the parasite to the cytoplasmic face of the erythrocyte membrane. The truncated PfEMP3 accumulates in structures that appear to be associated with the erythrocyte membrane. We show that accumulation of mutated PfEMP3 blocks the transfer of PfEMP1 onto the outside of the parasitized cell surface and suggest that these proteins traffic through an erythrocyte membrane-associated compartment that is involved in the transfer of PfEMP1 to the surface of the parasite-infected red blood cell. AD - Division of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research, Department of Pathology, The University of Melbourne, Melbourne 3050, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10856227 ID - 117 ER - TY - JOUR AU - Wang, L. AU - Menting, J. G. AU - Stowers, A. AU - Charoenvit, Y. AU - Sacci, J. B., Jr. AU - Coppel, R. L. PY - 2000 TI - Antigens cross reactive with Plasmodium falciparum merozoite surface protein 4 are found in pre-erythrocytic and sexual stages SP - 189-94 N1 - Jul JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 109 IS - 2 SN - 0166-6851 (Print) N1 - Antigens cross reactive with Plasmodium falciparum merozoite surface protein 4 are found in pre-erythrocytic and sexual stages N1 - 10960179 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Netherlands N1 - eng KW - Animals Antigens, Protozoan/genetics/immunology/*metabolism Base Sequence Cross Reactions Epitopes Fluorescent Antibody Technique, Indirect Humans Malaria, Falciparum/immunology/parasitology Molecular Sequence Data Plasmodium falciparum/genetics/growth & development/*immunology Protozoan Proteins/genetics/immunology/*metabolism Sequence Analysis, DNA AD - Department of Microbiology, Monash University, Vic., 3800, Clayton, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10960179 ID - 114 ER - TY - JOUR AU - Wang, L. AU - Menting, J. G. AU - Black, C. G. AU - Stowers, A. AU - Kaslow, D. C. AU - Hoffman, S. L. AU - Coppel, R. L. PY - 2000 TI - Differences in epitope recognition, isotype and titer of antisera to Plasmodium falciparum merozoite surface protein 4 raised by different modes of DNA or protein immunization SP - 816-24 N1 - Nov 22 JF - Vaccine JO - Vaccine VL - 19 IS - 7-8 SN - 0264-410X (Print) N1 - Differences in epitope recognition, isotype and titer of antisera to Plasmodium falciparum merozoite surface protein 4 raised by different modes of DNA or protein immunization N1 - 11115704 N1 - Dk32094/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Antibodies, Protozoan/biosynthesis Antigens, Protozoan/*genetics/*immunology COS Cells Epitopes/genetics Female Gene Expression Genes, Protozoan Humans Malaria Vaccines/genetics/*immunology Mice Mice, Inbred C3H Plasmodium falciparum/*genetics/*immunology Protozoan Proteins/*genetics/*immunology Transfection Vaccines, DNA/genetics/*immunology Vaccines, Subunit/genetics/immunology N2 - Plasmodium falciparum merozoite surface protein 4 (MSP4) is being developed as a component of a subunit vaccine against asexual stages of malaria. Three DNA constructs were produced that induced expression of MSP4 either in the cytoplasm of transfected cells or secreted from cells under the control of the human tissue plasminogen activator (TPA) signal or the native P. falciparum MSP4 signal. Only the construct containing the TPA signal induced detectable antibodies in mice, although gene expression was demonstrated in all constructs and MSP4 was shown to be secreted using either signal by in vitro transient transfection of COS cells. Two recombinant MSP4 proteins that encoded the same sequence as the plasmid DNA were produced in E. coli (EcMSP4-His) and S. cerevisiae (yMSP4-His) and used to raise antibodies in mice. Comparison of the antibodies elicited by these various antigen formulations showed differences in titer, isotype and epitope recognition. The titer of antibodies induced by DNA vaccination was lower than that induced by yMSP4-His, which in turn was lower than that induced by EcMSP4-His. The isotype profiles of the antibodies were also different, the plasmid DNA induced predominantly IgG(2a) responses whereas the two proteins induced predominantly IgG(1) responses. The antibodies induced by DNA and yMSP4-His recognized predominantly the C-terminal epidermal growth factor (EGF)-like domain of the protein, whereas EcMSP4-His induced antibodies recognizing all domains of the protein equally. The antibodies induced by DNA vaccination were directed almost extensively to conformational epitopes so that reactivity with native MSP4 was abolished after disulfide bonds in the protein were disrupted. Antibodies induced by recombinant proteins recognized linear epitopes as well and reactivity to native MSP4 was preserved after reduction and alkylation of parasite proteins. AD - Department of Microbiology, Monash University, Vic., 3800, Clayton, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11115704 ID - 108 ER - TY - JOUR AU - Tanaka, A. AU - Nalbandian, G. AU - Leung, P. S. AU - Benson, G. D. AU - Munoz, S. AU - Findor, J. A. AU - Branch, A. D. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2000 TI - Mucosal immunity and primary biliary cirrhosis: presence of antimitochondrial antibodies in urine SP - 910-5 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 32 IS - 5 SN - 0270-9139 (Print) N1 - Mucosal immunity and primary biliary cirrhosis: presence of antimitochondrial antibodies in urine N1 - 11050038 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Autoantibodies/blood/chemistry/classification/*urine Autoantigens/urine Epitope Mapping Female Humans *Immunity Immunoglobulin A/immunology/urine Immunoglobulin G/urine Liver Cirrhosis, Biliary/blood/*immunology/*urine Mitochondria/*immunology Protein Isoforms/urine Proteinuria/urine Urothelium/*immunology N2 - We have shown that IgA-class antimitochondrial autoantibodies (AMA) can be detected in the bile and saliva of patients with PBC, suggesting that AMA are secreted into the luminal fluid across bile ducts and salivary glands. These data prompted us to determine whether AMA of the IgA isotype may be transported across other epithelial mucosa. Therefore, we tested for the presence of AMA in the urine specimens of 83 patients with PBC and 58 non-PBC controls including healthy individuals and patients with other liver diseases. Patients enrolled in this study had no history of renal disease, and we confirmed there was less than 50 microgram/mL of protein in each of the urine specimens. Interestingly, we found that AMA were present in the urine of 71/83 (86%) of all patients with PBC and in 71/78 (91%) of patients with PBC that were serum AMA positive. In contrast, AMA were not detected in any of the 58 control urine specimens. Of particular interest, AMA of the IgA isotype was present in 57/83 (69%) of patients with PBC, and in 52 of these 57, we found secretory-type IgA. In a nested random subgroup of urine samples, the prevalence of the IgA2 AMA was 6/18 (33%), significantly lower than in matched serum samples, 13/16 (81%, P =.007). These data show that AMA of the IgA isotype is secreted into urine from the uroepithelium of patients with PBC, and support the thesis that PBC originated from either a mucosal challenge or a loss of mucosal tolerance. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Davis, California, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11050038 ID - 110 ER - TY - JOUR AU - Sasaki, M. AU - Ansari, A. A. AU - Nakanuma, Y. AU - Coppel, R. L. AU - Keeffe, E. B. AU - Gershwin, M. E. PY - 2000 TI - The immunopathology of primary biliary cirrhosis: thoughts for the millennium SP - 1-10 JF - Arch Immunol Ther Exp (Warsz) JO - Archivum immunologiae et therapiae experimentalis VL - 48 IS - 1 SN - 0004-069X (Print) N1 - The immunopathology of primary biliary cirrhosis: thoughts for the millennium N1 - 10722225 N1 - Journal Article Review Poland N1 - eng KW - Apoptosis Autoantibodies/metabolism Autoantigens Autoimmunity Cell Adhesion Molecules/metabolism Cytokines/metabolism Epithelium/immunology/pathology Female HLA Antigens/genetics Humans Inflammation/immunology/pathology Liver Cirrhosis, Biliary/genetics/*immunology/*pathology Male Mitochondria/metabolism Receptors, Antigen, T-Cell, alpha-beta/genetics T-Lymphocytes/immunology N2 - Primary biliary cirrhosis is an organ specific autoimmune disease that produces progressive cholestatic liver failure. It is predominantly a disease of women characterized by chronic progressive destruction of small intrahepatic bile ducts with portal inflammation and ultimately fibrosis. The serologic hallmark of primary biliary cirrhosis (PBC) is the presence of antibodies to mitochondria. The mechanisms by which and if which such antibodies produce liver tissue injury is unknown. However, the presence of these antibodies have allowed detailed immunological definition of the antigenic epitopes, the nature of reacting autoantibodies and the characterization of T cell responses. Several mechanisms may now be proposed regarding the immune mediated bile duct damage in PBC, including the possible role of T cell-mediated cytotoxicity and intracellular interaction between the IgA class of antimitochondrial antibodies (AMA) and mitochondrial autoantigens. The advent of molecular biology, the ability to clone and define epitopes, and the use of in situ nucleic acid hybridization, have all led to advances in understanding the natural history of immunopathology in PBC. There are major questions which remain unanswered, including, of course, etiology, but also including the questions of why there is female predominance, the absence of PBC in children, the relative ineffectiveness of immunosuppressive drugs, and the specific role of mitochondrial antigens. In this review, we focus on these issues and particularly on the immunobiology of patients with this disease. AD - Division of Allergy, Rheumatology, and Clinical Immunology, University of California, Davis, School of Medicine, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10722225 ID - 120 ER - TY - JOUR AU - Sasaki, M. AU - Ansari, A. AU - Pumford, N. AU - van de Water, J. AU - Leung, P. S. AU - Humphries, K. M. AU - Szweda, L. I. AU - Nakanuma, Y. AU - Roche, T. E. AU - Coppel, R. L. AU - Bach, J. F. AU - Gershwin, M. E. PY - 2000 TI - Comparative immunoreactivity of anti-trifluoroacetyl (TFA) antibody and anti-lipoic acid antibody in primary biliary cirrhosis: searching for a mimic SP - 51-60 N1 - Aug JF - J Autoimmun JO - Journal of autoimmunity VL - 15 IS - 1 SN - 0896-8411 (Print) N1 - Comparative immunoreactivity of anti-trifluoroacetyl (TFA) antibody and anti-lipoic acid antibody in primary biliary cirrhosis: searching for a mimic N1 - 10936028 N1 - Dk39588/dk/niddk Es10319/es/niehs Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Antigen-Antibody Reactions Autoantibodies/*chemistry/*metabolism Cattle Cytosol/drug effects/immunology/metabolism Dihydrolipoyllysine-Residue Acetyltransferase Enzyme Inhibitors/immunology Enzyme-Linked Immunosorbent Assay Halothane/administration & dosage Haptens/immunology Hemocyanin/immunology Humans Immune Sera/metabolism Immunoblotting Immunohistochemistry Liver Cirrhosis, Biliary/blood/enzymology/*immunology Microsomes, Liver/drug effects/immunology/metabolism Molecular Mimicry/*immunology Mollusca Pyruvate Dehydrogenase Complex/antagonists & inhibitors/metabolism Rats Serum Albumin/immunology Thioctic Acid/*immunology Trifluoroacetic Acid/*analogs & derivatives/*immunology N2 - Previous studies documenting the existence of cross-reactivity between the lipoated (but not unlipoated) forms of the inner lipoyl domain (E2L2) of PDC-E2 [the major autoantigen in Primary biliary cirrhosis (PBC)] and trifluoroacetylated (TFA) proteins, led us to hypothesize that PBC may be due to an initial insult with an environmental agent that cross-reacts with TFA. Therefore, we performed a comparative study of the reactivity of rabbit anti-TFA antibody and anti-lipoic acid (LA) antibody against the mitochondrial autoantigens of human PBC and various TFA and LA conjugated proteins. Whereas both anti-TFA and anti-LA reacted with PDC-E2, the wild-type lipoated form of E2L2, OGDC-E2, E3-BP and LA-KLH, neither reacted with BCOADC-E2 or the non-lipoated form of E2L2. Of interest was that while anti-TFA reacted with PDC-E2, TFA-RSA and LA-KLH, it failed to inhibit PDC-E2 enzyme function. In contrast, anti-LA demonstrated cytoplasmic and mitochondrial staining, and inhibited PDC enzyme activity. Hence, although considerable cross reactivity exists between anti-TFA and anti-LA, the molecular nature of the interaction is clearly different. One of 14 PBC sera reacted weakly with TFA-albumin, whereas four of 14 PBC sera reacted with LA-KLH. Immunohistochemically, both anti-TFA and anti-LA antibodies reacted focally with periportal hepatocytes and bile ducts in both PBC and controls. However, anti-LA produced much stronger focalized staining of the bile ducts of diseased liver. This study suggests that while anti-TFA antibody recognizes lipoic acid-linked enzymes and proteins, the epitope recognized differs from that of anti-LA antibody and PBC autoantibodies. It is unlikely that a response to TFA is the triggering event in PBC. Anti-LA antibodies share a higher degree of similarity to PBC sera providing suggestive evidence that anti-LA antibodies or anti-LA like antibodies (mimotopes) may help define the initiator of the autoimmune response. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10936028 ID - 116 ER - TY - JOUR AU - Reynoso-Paz, S. AU - Leung, P. S. AU - Van De Water, J. AU - Tanaka, A. AU - Munoz, S. AU - Bass, N. AU - Lindor, K. AU - Donald, P. J. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 2000 TI - Evidence for a locally driven mucosal response and the presence of mitochondrial antigens in saliva in primary biliary cirrhosis SP - 24-9 N1 - Jan JF - Hepatology JO - Hepatology (Baltimore, Md VL - 31 IS - 1 SN - 0270-9139 (Print) N1 - Evidence for a locally driven mucosal response and the presence of mitochondrial antigens in saliva in primary biliary cirrhosis N1 - 10613723 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Antibody Specificity Autoantibodies/*analysis/blood Autoantigens/*immunology Epitope Mapping Humans Immunoglobulin A/analysis Immunoglobulin G/analysis Immunoglobulin M/analysis Liver Cirrhosis, Biliary/*immunology Mitochondria/*immunology Mouth Mucosa/*immunology Pyruvate Dehydrogenase Complex/immunology Saliva/*immunology N2 - Primary biliary cirrhosis (PBC) is often considered to be a dry gland disease caused by frequent involvement of salivary and lacrimal glands. Although high titers of antimitochondrial autoantibodies (AMA) have long been recognized in PBC, little is known about the presence of mitochondrial autoantigens in mucosal compartments such as saliva. We investigated saliva and sera in PBC patients and controls for the presence of AMA and mitochondrial antigens. In PBC saliva, AMA were detected in 45 of 49 (92%), with specificity directed against pyruvate dehydrogenase complex (PDC-E2) alone in 22 of 49 (45%), against PDC-E2 and branched-chain 2-oxo-acid dehydrogenase complex E2 (BCOADC-E2) in 4 of 49 (8%), to PDC-E2 and 2-oxoglutarate dehydrogenase complex E2 (OGDC-E2) in 9 of 49 (18%), and to the 3 antigens together in 10 of 49 (20%). Isotyping of the saliva AMA showed that 80% of the patients had immunoglobulin A (IgA) against PDC-E2, 18% had IgM-specific PDC-E2, and 35% had IgG specific PDC-E2. Similar to serum and bile anti-PDC-E2 IgA antibodies, the saliva autoantibodies localized their reactivity to the inner lipoyl domain of PDC-E2. Furthermore, saliva from patients with PBC but not controls inhibited pyruvate dehydrogenase enzyme activity in vitro. In addition, and of particular interest, we detected a molecule with a molecular weight corresponding to PDC-E2 (74 kd) in PBC but not control saliva. These findings make several important points: first, there appears to be localized mucosal immunity in the secretory system of PBC; second, AMA are readily detected in PBC saliva; and third, PDC-E2 may be present in the saliva of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10613723 ID - 122 ER - TY - JOUR AU - Reynoso-Paz, S. AU - Coppel, R. L. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 2000 TI - Primary biliary cirrhosis. Connecting molecular biology to clinical medicine SP - 241-62 N1 - Apr JF - Clin Rev Allergy Immunol JO - Clinical reviews in allergy & immunology VL - 18 IS - 2 SN - 1080-0549 (Print) N1 - Primary biliary cirrhosis. Connecting molecular biology to clinical medicine N1 - 10944707 N1 - Journal Article Review United states N1 - eng KW - Autoantibodies/analysis/immunology Autoantigens/genetics/immunology *Autoimmune Diseases/immunology/pathology/physiopathology Bile Ducts, Intrahepatic/immunology Cytokines/immunology Female Granuloma/pathology Humans *Liver Cirrhosis, Biliary/immunology/pathology/physiopathology Male Mitochondria, Liver/immunology Molecular Mimicry T-Lymphocytes/immunology AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, School of Medicine 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10944707 ID - 115 ER - TY - JOUR AU - Patterson, J. H. AU - McConville, M. J. AU - Haites, R. E. AU - Coppel, R. L. AU - Billman-Jacobe, H. PY - 2000 TI - Identification of a methyltransferase from Mycobacterium smegmatis involved in glycopeptidolipid synthesis SP - 24900-6 N1 - Aug 11 JF - J Biol Chem JO - The Journal of biological chemistry VL - 275 IS - 32 SN - 0021-9258 (Print) N1 - Identification of a methyltransferase from Mycobacterium smegmatis involved in glycopeptidolipid synthesis N1 - 10801784 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Cell Wall/chemistry Conserved Sequence Glycoconjugates/*biosynthesis/chemistry/isolation & purification Methylation Methyltransferases/chemistry/*genetics/*metabolism Molecular Sequence Data Multigene Family Mycobacterium smegmatis/*enzymology/*genetics Open Reading Frames Rhamnose/metabolism S-Adenosylmethionine/metabolism Sequence Alignment Sequence Homology, Amino Acid N2 - Glycopeptidolipids (GPLs) are major components of the cell walls of several species of mycobacteria. We have isolated a transposon mutant of Mycobacterium smegmatis that is unable to synthesize mature GPLs and that displays a rough colony morphology. The disrupted gene, mtf1, shares a high degree of homology with several S-adenosylmethionine-dependent methyltransferases. The enzyme encoded by mtf1 is required for the methylation of a single rhamnose residue that forms part of the conserved GPL core structure. This conclusion is supported by the finding that (a) the mutant synthesized only GPLs with undermethylated (either mono- or nonmethylated instead of di- or trimethylated) rhamnose residues; (b) complementation of the mutant with a wild-type copy of mtf1 restored high levels of synthesis of GPLs containing di- and trimethylated rhamnose; and (c) S-adenosylmethionine-dependent methylation of rhamnosylated GPLs could be detected in cell lysates of wild-type cells and mtf1-complemented mutant cells, but not in mutant cells lacking intact mtf1. Structural analysis of wild-type and mutant GPLs suggests that disruption of mtf1 specifically inhibits addition of O-methyl groups to the 3 (or 2)-position of the rhamnose. In the absence of 3-O-methylation, further methylation of GPL rhamnose is apparently inhibited, and overall GPL synthesis is down-regulated by 90%. AD - Department of Biochemistry and Molecular Biology, University of Melbourne, Royal Parade, Parkville, Victoria 3052, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10801784 ID - 119 ER - TY - JOUR AU - Nishio, A. AU - Coppel, R. AU - Ishibashi, H. AU - Gershwin, M. E. PY - 2000 TI - The pyruvate dehydrogenase complex as a target autoantigen in primary biliary cirrhosis SP - 535-47 N1 - Aug JF - Baillieres Best Pract Res Clin Gastroenterol JO - Bailliere's best practice & research VL - 14 IS - 4 SN - 1521-6918 (Print) N1 - The pyruvate dehydrogenase complex as a target autoantigen in primary biliary cirrhosis N1 - 10976013 N1 - Journal Article Review England N1 - eng KW - Autoantigens/*immunology Autoimmune Diseases/*immunology B-Lymphocytes/immunology Bile/immunology Dihydrolipoyllysine-Residue Acetyltransferase Epitopes/immunology Female Humans Liver Cirrhosis, Biliary/*immunology Male Pyruvate Dehydrogenase Complex/*immunology T-Lymphocytes/immunology N2 - Mitochondrial autoantigens and their B and T cell autoepitopes have been well defined in primary biliary cirrhosis (PBC). However, the relationships of the antimitochondrial antibodies and the mechanisms of bile duct destruction in PBC remain an enigma. The serological hallmark of PBC remains the presence of antibodies to mitochondria, particularly to the E2 component of the pyruvate dehydrogenase complex (PDC-E2). However, several mechanisms may now be proposed which may explain the immune-mediated bile duct damage in PBC. These include the possible role of T cell-mediated cytotoxicity as well as the interaction between the IgA class of antimitochondrial antibodies and the mitochondrial autoantigens. A prominent feature in this discussion is the highly directed and specific immune response to the mitochondrial antigens, including PDC-E2 as well as other members of the 2-oxo-acid dehydrogenase complexes. Ultimately, the mechanisms that lead to this immune reaction should provide data on other questions in PBC, including the reasons for female predominance, the absence of PBC in children and the relative ineffectiveness of immunosuppressive agents. AD - Department of Gastroenterology, Tenri Hospital, Nara, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10976013 ID - 113 ER - TY - JOUR AU - Magowan, C. AU - Nunomura, W. AU - Waller, K. L. AU - Yeung, J. AU - Liang, J. AU - Van Dort, H. AU - Low, P. S. AU - Coppel, R. L. AU - Mohandas, N. PY - 2000 TI - Plasmodium falciparum histidine-rich protein 1 associates with the band 3 binding domain of ankyrin in the infected red cell membrane SP - 461-70 N1 - Nov 15 JF - Biochim Biophys Acta JO - Biochimica et biophysica acta VL - 1502 IS - 3 SN - 0006-3002 (Print) N1 - Plasmodium falciparum histidine-rich protein 1 associates with the band 3 binding domain of ankyrin in the infected red cell membrane N1 - 11068188 N1 - Dk32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals Anion Exchange Protein 1, Erythrocyte/*metabolism Ankyrins/chemistry/*metabolism Binding Sites Erythrocyte Membrane/*metabolism Malaria, Falciparum/*blood Peptides/chemistry/*metabolism Phosphoproteins/metabolism *Plasmodium falciparum Protozoan Proteins/*metabolism N2 - Infection of erythrocytes by the malaria parasite Plasmodium falciparum results in the export of several parasite proteins into the erythrocyte cytoplasm. Changes occur in the infected erythrocyte due to altered phosphorylation of proteins and to novel interactions between host and parasite proteins, particularly at the membrane skeleton. In erythrocytes, the spectrin based red cell membrane skeleton is linked to the erythrocyte plasma membrane through interactions of ankyrin with spectrin and band 3. Here we report an association between the P. falciparum histidine-rich protein (PfHRP1) and phosphorylated proteolytic fragments of red cell ankyrin. Immunochemical, biochemical and biophysical studies indicate that the 89 kDa band 3 binding domain and the 62 kDa spectrin-binding domain of ankyrin are co-precipitated by mAb 89 against PfHRP1, and that native and recombinant ankyrin fragments bind to the 5' repeat region of PfHRP1. PfHRP1 is responsible for anchoring the parasite cytoadherence ligand to the erythrocyte membrane skeleton, and this additional interaction with ankyrin would strengthen the ability of PfEMP1 to resist shear stress. AD - Lawrence Berkeley National Laboratory Life Sciences Division, Berkeley, CA 94720, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11068188 ID - 109 ER - TY - JOUR AU - Kedzierski, L. AU - Black, C. G. AU - Coppel, R. L. PY - 2000 TI - Characterization of the merozoite surface protein 4/5 gene of Plasmodium berghei and Plasmodium yoelii SP - 137-47 N1 - Jan 5 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 105 IS - 1 SN - 0166-6851 (Print) N1 - Characterization of the merozoite surface protein 4/5 gene of Plasmodium berghei and Plasmodium yoelii N1 - 10613706 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Motifs Amino Acid Sequence Animals Antibodies, Protozoan/blood/immunology Antigens, Protozoan/chemistry/*genetics/immunology Cloning, Molecular Cross Reactions DNA, Protozoan/analysis/genetics Female *Genes, Protozoan Immunoblotting Malaria/parasitology Membrane Proteins/chemistry/*genetics/immunology Mice Mice, Inbred BALB C Molecular Sequence Data Plasmodium berghei/*genetics/immunology Plasmodium yoelii/*genetics/immunology Protozoan Proteins/chemistry/*genetics/immunology Recombinant Proteins/immunology/metabolism Reverse Transcriptase Polymerase Chain Reaction N2 - The genes encoding merozoite surface protein 4/5 (MSP4/5) from Plasmodium berghei and Plasmodium yoelii have been cloned and completely sequenced. Comparisons of the predicted protein sequences with those of Plasmodium chabaudi MSP4/5 and Plasmodium falciparum MSP4 and MSP5 show general structural similarities. All predicted proteins contain hydrophobic signal sequences, potential GPI attachment sequences and a single epidermal growth factor (EGF)-like domain at the C-terminus. The amino acid sequence of the EGF-like motif is highly conserved in rodent malaria species and also shows a considerable degree of similarity with the EGF-like domains found in the P. falciparum proteins. Both the P. yoelii and P. berghei genes show evidence of both spliced and unspliced mRNA at steady state. This phenomenon is similar to that seen for the P. chabaudi MSP4/5 gene, and is believed to be involved in regulation of protein expression. We describe here the construction of clones expressing full length recombinant protein. Antibodies directed against recombinant MSP4/5 proteins recognize a single polypeptide on parasite material and show crossreactivity between MSP4/5 from different murine malaria species, but do not crossreact with either MSP4 or MSP5 from P. falciparum. The various antisera show reactivity against reduction sensitive epitopes as well as reduction insensitive epitopes. AD - Department of Microbiology, Monash University, Clayton, Vic., Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10613706 ID - 123 ER - TY - JOUR AU - Kedzierski, L. AU - Black, C. G. AU - Coppel, R. L. PY - 2000 TI - Immunization with recombinant Plasmodium yoelii merozoite surface protein 4/5 protects mice against lethal challenge SP - 6034-7 N1 - Oct JF - Infect Immun JO - Infection and immunity VL - 68 IS - 10 SN - 0019-9567 (Print) N1 - Immunization with recombinant Plasmodium yoelii merozoite surface protein 4/5 protects mice against lethal challenge N1 - 10992516 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antibodies, Protozoan/blood Antigens, Protozoan/administration & dosage/genetics/*immunology Female Malaria/*prevention & control *Malaria Vaccines/administration & dosage/immunology Mice Mice, Inbred BALB C Parasitemia/parasitology Plasmodium yoelii/*immunology Protozoan Proteins/administration & dosage/genetics/*immunology Recombinant Proteins/administration & dosage/immunology Vaccination N2 - Plasmodium yoelii merozoite surface protein 4/5 (PyMSP4/5), expressed as a recombinant protein, was highly effective at protecting mice against lethal challenge with P. yoelii. There was a significant correlation between prechallenge antibody levels and peak parasitemia, suggesting that the homologues of PyMSP4/5 in Plasmodium falciparum are promising components of a subunit vaccine against malaria. AD - Department of Microbiology, Monash University 3800, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10992516 ID - 112 ER - TY - JOUR AU - Gershwin, M. E. AU - Ansari, A. A. AU - Mackay, I. R. AU - Nakanuma, Y. AU - Nishio, A. AU - Rowley, M. J. AU - Coppel, R. L. PY - 2000 TI - Primary biliary cirrhosis: an orchestrated immune response against epithelial cells SP - 210-25 N1 - Apr JF - Immunol Rev JO - Immunological reviews VL - 174 SN - 0105-2896 (Print) N1 - Primary biliary cirrhosis: an orchestrated immune response against epithelial cells N1 - 10807518 N1 - Comparative Study Journal Article Review Denmark N1 - eng KW - Acyltransferases/*immunology Age Distribution Amino Acid Motifs Amino Acid Sequence Animals Autoantibodies/*immunology Autoantigens/*immunology Autoimmune Diseases/epidemiology/*immunology Bacterial Proteins/immunology Bile Ducts/immunology/pathology Cell Line Cross Reactions Cytokines/physiology Dihydrolipoyllysine-Residue Acetyltransferase Dogs Epithelial Cells/immunology/pathology Escherichia coli Infections/complications/immunology Female Genetic Predisposition to Disease HLA-D Antigens/genetics/immunology Humans Immunodominant Epitopes/immunology Immunoglobulin A/immunology Liver Cirrhosis, Biliary/epidemiology/etiology/*immunology Male Maternal-Fetal Exchange Middle Aged Mitochondria, Liver/*immunology Molecular Mimicry Molecular Sequence Data Peptides/*immunology Pregnancy Protein Structure, Tertiary Pyruvate Dehydrogenase Complex/*immunology Scleroderma, Systemic/immunology/pathology Sequence Alignment Sequence Homology, Amino Acid Sex Distribution N2 - Primary biliary cirrhosis (PBC) is an organ-specific autoimmune disease that predominantly affects women and is characterized by chronic progressive destruction of small intrahepatic bile ducts with portal inflammation and ultimately fibrosis. The serologic hallmark of PBC is the presence of antibodies to mitochondria, especially to the E2 component of the pyruvate dehydrogenase complex. The mechanisms by which (and if) such antibodies produce liver tissue injury are unknown. However, the presence of these antibodies has allowed detailed immunological definition of the antigenic epitopes, the nature of reactive autoantibodies and the characterization of T-cell responses. Several mechanisms may now be proposed regarding the immune-mediated bile duct damage in PBC, including the possible role of T-cell-mediated cytotoxicity and intracellular interaction between the IgA class of antimitochondrial antibodies and mitochondrial autoantigens. There are major questions which remain unanswered, including, of course, etiology, but also the reasons for female predominance, the absence of PBC in children, the relative ineffectiveness of immunosuppressive drugs, and the specific role of mitochondrial antigens. The data so far provide suggestive evidence that PBC is a mucosal disease; this thesis provides a basis for discussion of etiology via the enterohepatic circulation of toxins and/or infection. AD - Division of Allergy, Rheumatology, and Clinical Immunology, University of California, Davis, School of Medicine, 95616, USA. megershwin@ucdavis.edu UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10807518 ID - 118 ER - TY - JOUR AU - Cooke, B. AU - Coppel, R. AU - Wahlgren, M. PY - 2000 TI - Falciparum malaria: sticking up, standing out and out-standing SP - 416-20 N1 - Oct JF - Parasitol Today JO - Parasitology today (Personal ed VL - 16 IS - 10 SN - 0169-4758 (Print) N1 - Falciparum malaria: sticking up, standing out and out-standing N1 - 11006472 N1 - Journal Article Review England N1 - eng KW - Animals Cell Adhesion Erythrocytes/*parasitology Humans Malaria, Falciparum/*parasitology/physiopathology Plasmodium falciparum/genetics/*pathogenicity/physiology Virulence N2 - Cytoadherence is believed to be fundamental for the survival of Plasmodium falciparum in vivo and, uniquely, is a major determinant of the virulence of this parasite. Despite the widely professed importance of cytoadhesion in the development of severe disease, there are a number of aspects of this highly complex process that remain poorly understood. Recent progress in the understanding of cytoadhesive phenomena was discussed extensively at the Molecular Approaches to Malaria conference, Lorne, Australia, 2-5 February 2000. Here, Brian Cooke, Mats Wahlgren and Ross Coppel consider just how far we have progressed during the past 30 years and highlight what is still missing in our understanding of the mechanisms and clinical relevance of this apparently vital process. AD - Department of Microbiology, PO Box 53, Monash University, 3800, Victoria, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11006472 ID - 111 ER - TY - JOUR AU - Black, C. G. AU - Coppel, R. L. PY - 2000 TI - Synonymous and non-synonymous mutations in a region of the Plasmodium chabaudi genome and evidence for selection acting on a malaria vaccine candidate SP - 447-51 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 111 IS - 2 SN - 0166-6851 (Print) N1 - Synonymous and non-synonymous mutations in a region of the Plasmodium chabaudi genome and evidence for selection acting on a malaria vaccine candidate N1 - 11163451 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Adenylosuccinate Lyase/genetics Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence Evolution, Molecular Genes, Protozoan *Genome, Protozoan Humans Malaria/prevention & control Malaria Vaccines Membrane Proteins/*genetics Molecular Sequence Data *Mutation Plasmodium chabaudi/*genetics/immunology Protozoan Proteins/*genetics AD - Department of Microbiology, Monash University, Vic, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11163451 ID - 106 ER - TY - JOUR AU - Wu, T. AU - Black, C. G. AU - Wang, L. AU - Hibbs, A. R. AU - Coppel, R. L. PY - 1999 TI - Lack of sequence diversity in the gene encoding merozoite surface protein 5 of Plasmodium falciparum SP - 243-50 N1 - Oct 15 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 103 IS - 2 SN - 0166-6851 (Print) N1 - Lack of sequence diversity in the gene encoding merozoite surface protein 5 of Plasmodium falciparum N1 - 10551366 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan *Antigenic Variation Base Sequence *Genes, Protozoan Introns Membrane Proteins/*genetics/immunology Molecular Sequence Data Plasmodium falciparum/*genetics Polymerase Chain Reaction Sequence Analysis, DNA Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid N2 - The gene encoding merozoite surface protein 5 (MSP5) of Plasmodium falciparum is situated between the genes encoding MSP2 and MSP4 on chromosome 2. Both MSP4 and MSP5 encode proteins that contain hydrophobic signal and glycosylphosphatidylinositol (GPI) attachment signals and a single epidermal growth factor (EGF)-like domain at their carboxyl termini. The similar gene organization, location and similar structural features of the two genes suggest that they have arisen from a gene duplication event. In this study we provide further evidence for the merozoite surface location of MSP5 by demonstrating that MSP5 is present in isolated merozoites, partitions in the detergent-enriched phase following Triton X-114 fractionation and shows a staining pattern consistent with merozoite surface location by indirect immunofluorescence confocal microscopy. Analysis of antigenic diversity of MSP5 shows a lack of sequence variation between various isolates of P. falciparum from different geographical locations, a feature unusual for surface proteins of merozoites and one that may simplify vaccine formulation. AD - Department of Microbiology, Monash University, Clayton, Vic., Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10551366 ID - 126 ER - TY - JOUR AU - Wang, L. AU - Black, C. G. AU - Marshall, V. M. AU - Coppel, R. L. PY - 1999 TI - Structural and antigenic properties of merozoite surface protein 4 of Plasmodium falciparum SP - 2193-200 N1 - May JF - Infect Immun JO - Infection and immunity VL - 67 IS - 5 SN - 0019-9567 (Print) N1 - Structural and antigenic properties of merozoite surface protein 4 of Plasmodium falciparum N1 - 10225874 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United states N1 - eng KW - Animals Antibodies, Protozoan/blood Antigens, Protozoan/*chemistry/genetics Antigens, Surface/chemistry/genetics Epitope Mapping Humans Immunization Malaria, Falciparum/immunology/parasitology Peptide Fragments/chemistry/genetics/immunology Plasmodium falciparum/genetics/growth & development/*immunology Protein Conformation Protozoan Proteins/*chemistry/genetics/*immunology Rabbits Recombinant Fusion Proteins/chemistry/genetics/immunology N2 - Merozoite surface protein 4 (MSP4) of Plasmodium falciparum is a glycosylphosphatidylinositol-anchored integral membrane protein of 272 residues that possesses a single epidermal growth factor (EGF)-like domain near the carboxyl terminus. We have expressed both full-length MSP4 and a number of fragments in Escherichia coli and have used these recombinant proteins to raise experimental antisera. All recombinant proteins elicited specific antibodies that reacted with parasite-derived MSP4 by immunoblotting. Antibody reactivity was highly dependent on the protein conformation. For example, reduction and alkylation of MSP4 almost completely abolished the reactivity of several antibody preparations, including specificities directed to regions of the protein that do not contain cysteine residues and are far removed from the cysteine-containing EGF-like domain. This indicated the presence of conformation-dependent epitopes in MSP4 and demonstrated that proper folding of the EGF-like domain influenced the antigenicity of the entire molecule. The recombinant proteins were used to map epitopes recognized by individuals living in areas where malaria is endemic, and at least four distinct regions are naturally antigenic during infection. Binding of human antibodies to the EGF-like domain was essentially abrogated after reduction of the recombinant protein, indicating the recognition of conformational epitopes by the human immune responses. This observation led us to examine the importance of conformation dependence in responses to other integral membrane proteins of asexual stages. We analyzed the natural immune responses to a subset of these antigens and demonstrated that there is diminished reactivity to several antigens after reduction. These studies demonstrate the importance of reduction-sensitive structures in the maintenance of the antigenicity of several asexual-stage antigens and in particular the importance of the EGF-like domain in the antigenicity of MSP4. AD - Department of Microbiology, Monash University, Clayton, Victoria, 3168, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10225874 ID - 135 ER - TY - JOUR AU - Waller, K. L. AU - Cooke, B. M. AU - Nunomura, W. AU - Mohandas, N. AU - Coppel, R. L. PY - 1999 TI - Mapping the binding domains involved in the interaction between the Plasmodium falciparum knob-associated histidine-rich protein (KAHRP) and the cytoadherence ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1) SP - 23808-13 N1 - Aug 20 JF - J Biol Chem JO - The Journal of biological chemistry VL - 274 IS - 34 SN - 0021-9258 (Print) N1 - Mapping the binding domains involved in the interaction between the Plasmodium falciparum knob-associated histidine-rich protein (KAHRP) and the cytoadherence ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1) N1 - 10446142 N1 - Dk-32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Base Sequence Binding Sites Merozoite Surface Protein 1/*chemistry Molecular Sequence Data Peptides/*chemistry Plasmodium falciparum/*chemistry Polymerase Chain Reaction Protozoan Proteins Repetitive Sequences, Amino Acid N2 - Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) clusters at electron-dense knob-like structures on the surface of malaria-infected red blood cells and mediates their adhesion to the vascular endothelium. In parasites lacking knobs, vascular adhesion is less efficient, and infected red cells are not able to immobilize successfully under hemodynamic flow conditions even though PfEMP1 is still present on the exterior of the infected red cell. We examined the interaction between the knob-associated histidine-rich protein (KAHRP), the parasite protein upon which knob formation is dependent, and PfEMP1, and we show evidence of a direct interaction between KAHRP and the cytoplasmic region of PfEMP1 (VARC). We have identified three fragments of KAHRP which bind VARC. Two of these KAHRP fragments (K1A and K2A) interact with VARC with binding affinities (K(D(kin))) of 1 x 10(-7) M and 3.3 x 10(-6) M respectively, values comparable to those reported previously for protein-protein interactions in normal and infected red cells. Further experiments localized the high affinity binding regions of KAHRP to the 63-residue histidine-rich and 70-residue 5' repeats. Deletion of these two regions from the KAHRP fragments abolished their ability to bind to VARC. Identification of the critical domains involved in interaction between KAHRP and PfEMP1 may aid development of new therapies to prevent serious complications of P. falciparum malaria. AD - Department of Microbiology, Monash University, Clayton, Victoria 3168, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10446142 ID - 130 ER - TY - JOUR AU - Tsuji, K. AU - Watanabe, Y. AU - Van De Water, J. AU - Nakanishi, T. AU - Kajiyama, G. AU - Parikh-Patel, A. AU - Coppel, R. AU - Gershwin, M. E. PY - 1999 TI - Familial primary biliary cirrhosis in Hiroshima SP - 171-8 N1 - Aug JF - J Autoimmun JO - Journal of autoimmunity VL - 13 IS - 1 SN - 0896-8411 (Print) N1 - Familial primary biliary cirrhosis in Hiroshima N1 - 10441183 N1 - Journal Article Review England N1 - eng KW - Adult Age of Onset Aged Animals Autoantibodies/blood Cattle Female HLA Antigens/genetics Humans Japan/epidemiology Liver Cirrhosis, Biliary/epidemiology/*genetics/*immunology Male Middle Aged Mitochondria/immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the presence of anti-mitochondrial antibodies and chronic inflammatory destruction of septal and intrahepatic bile ducts. Although there are no obvious associations of PBC with MHC class I or class II genes, there appears to be a significant increased risk of developing disease within families. Clearly, a combination of genetic and environmental factors play a role in disease pathogenesis, although the relative contributions of each are unclear. In this study, we have taken advantage of the well-defined health-care system in Hiroshima prefecture, where PBC is a reportable disease. In the period 1988-1997, 156 new patients with PBC in a total population of 2,873,000 were diagnosed. These patients included 18 subjects that were derived from eight different families in which more than one family member had a history of PBC; this reflects a frequency of 5.1% and further shows that the prevalence of PBC is greatly increased in family members. Of interest, the median age of onset of PBC in second generation patients was much younger (33.4+/-10.8 years) compared to median disease onset in general patients with PBC in Hiroshima (55.6+/-12 years). In fact, it was striking that the onset of disease in family members often occurred within a few years of each other. We also noted that sera of affected members had similar AMA reactive profiles against recombinant PDC-E2, BCKD-E2 and OGDC-E2; the major autoantigens of PBC. Similar HLA types were found within affected members of a pedigree but the data is limited because of absence of similar typing of unaffected members. The increased family history of PBC, and the earlier onset of disease in second generation members, suggests that environmental agents are an important risk factor for the development of disease. We suggest that genomic analysis in familial PBC will be important to identify the mechanisms of genetic susceptibility. AD - First Department of Internal Medicine, Hiroshima University School of Medicine, Hiroshima, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10441183 ID - 131 ER - TY - JOUR AU - Tanaka, A. AU - Quaranta, S. AU - Mattalia, A. AU - Coppel, R. AU - Rosina, F. AU - Manns, M. AU - Gershwin, M. E. PY - 1999 TI - The tumor necrosis factor-alpha promoter correlates with progression of primary biliary cirrhosis SP - 826-9 N1 - May JF - J Hepatol JO - Journal of hepatology VL - 30 IS - 5 SN - 0168-8278 (Print) N1 - The tumor necrosis factor-alpha promoter correlates with progression of primary biliary cirrhosis N1 - 10365808 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Denmark N1 - eng KW - Alleles California DNA/blood Disease Progression European Continental Ancestry Group Genotype Germany Heterozygote Detection Homozygote Humans Italy Liver Cirrhosis, Biliary/*genetics/*physiopathology Odds Ratio Polymerase Chain Reaction *Polymorphism, Restriction Fragment Length *Promoter Regions (Genetics) Reference Values Tumor Necrosis Factor-alpha/*genetics Victoria N2 - BACKGROUND/AIMS: There have been many studies attempting to identify genes that determine susceptibility to primary biliary cirrhosis (PBC), but few studies have attempted to define the genes that modulate the natural history of the disease. There is a biallelic polymorphism, coined TNF1 and TNF2, in the TNFalpha promoter region at -308. We investigated the relative frequency of the TNF1 and TNF2 alleles in patients with PBC, based on the hypothesis that a polymorphism of the TNFalpha promoter region may be associated with the rate of progression and prognosis of PBC. METHODS: Seventy-one Caucasoid patients with PBC and 133 healthy and unrelated Caucasoid individuals were studied. Genomic DNA was extracted from blood, and the mutation at position -308 of the TNFalpha gene analyzed by PCR and NcoI digestion. RESULTS: In 71 patients with PBC, 56/71 (78.9%) patients were TNF1/TNF1 homozygotes, 14/71 (19.7%) were TNF1/TNF2 heterozygotes and 1/71 (1.4%) were TNF2/TNF2 homozygotes. In 133 healthy individuals, 109/133 (80.5%) patients were TNF1/TNF1 homozygotes, 24/133 (18%) were TNF1/TNF2 heterozygotes. No control individuals were TNF2/TNF2 homozygotes. The difference between the two groups was not statistically significant (p = 0.3684). However, in patients with TNF1/TNF1 the Mayo score for disease severity was 4.596+/-0.157 (mean +/- SEM), compared to 5.637+/-0.420 for patients with TNF1/TNF2. This Mayo score was significantly higher in patients with the TNF1/TNF2 genotype than those with TNF1/TNF1 (p = 0.0140), with an odds ratio of 4.9. CONCLUSIONS: Our data demonstrate that the presence of the TNF2 allele may be associated with a higher Mayo score, and thus with patients in a more advanced clinical stage. These data have both theoretical and clinical implications. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10365808 ID - 134 ER - TY - JOUR AU - Tanaka, A. AU - Prindiville, T. P. AU - Gish, R. AU - Solnick, J. V. AU - Coppel, R. L. AU - Keeffe, E. B. AU - Ansari, A. AU - Gershwin, M. E. PY - 1999 TI - Are infectious agents involved in primary biliary cirrhosis? A PCR approach SP - 664-71 N1 - Oct JF - J Hepatol JO - Journal of hepatology VL - 31 IS - 4 SN - 0168-8278 (Print) N1 - Are infectious agents involved in primary biliary cirrhosis? A PCR approach N1 - 10551390 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Denmark N1 - eng KW - Archaea/genetics/isolation & purification Consensus Sequence Eubacterium/classification/genetics/isolation & purification Glyceraldehyde-3-Phosphate Dehydrogenases/genetics Helicobacter/genetics/isolation & purification Humans Liver/microbiology Liver Cirrhosis, Biliary/genetics/*microbiology Mycobacterium/genetics/isolation & purification Polymerase Chain Reaction Reference Values N2 - BACKGROUND/AIMS: A variety of data suggest that microbial infections and, in particular, atypical mycobacteria infections, may either initiate and/or be associated with the pathogenesis of primary biliary cirrhosis. METHODS: To address this hypothesis, use was made of polymerase chain reaction techniques and primers specific for the 16s rRNA gene of Eubacteria, Archaeabacteria, Mycobacteria and Helicobacter to determine if such sequences were detectable in liver tissue specimens from 29 patients with primary biliary cirrhosis. Similar liver tissues from patients with primary sclerosing cholangitis, chronic hepatitis, alcoholic liver disease and otherwise normal donors were analyzed in parallel. Genomic DNA was extracted from each of these liver tissue specimens using sterile techniques to avoid possible laboratory contamination. The DNA was subjected to polymerase chain reaction amplification using bacterial genus specific primers and the amplified products cloned and sequenced. Sequence data were analyzed by searching for homology to existing genes. RESULTS: Sequences from primary biliary cirrhosis and control livers corresponded to those found in a variety of bacteria, but no consensus sequence was found in primary biliary cirrhosis specimens. Neither Archaeabacteria nor Mycobacteria products were detected in liver specimens of patients with primary biliary cirrhosis, and Helicobacter pylori DNA was detected in only one primary biliary cirrhosis patient. CONCLUSIONS: Although bacterial infection, particularly with intracellular organisms, has been suggested to play a role in the initiation of primary biliary cirrhosis, there is no evidence from this study to suggest an ongoing chronic infectious process. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10551390 ID - 125 ER - TY - JOUR AU - Tanaka, A. AU - Lindor, K. AU - Gish, R. AU - Batts, K. AU - Shiratori, Y. AU - Omata, M. AU - Nelson, J. L. AU - Ansari, A. AU - Coppel, R. AU - Newsome, M. AU - Gershwin, M. E. PY - 1999 TI - Fetal microchimerism alone does not contribute to the induction of primary biliary cirrhosis SP - 833-8 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 30 IS - 4 SN - 0270-9139 (Print) N1 - Fetal microchimerism alone does not contribute to the induction of primary biliary cirrhosis N1 - 10498630 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged *Chimera Female Fetus/*physiology Humans Liver/physiopathology Liver Cirrhosis, Biliary/*genetics Middle Aged Polymerase Chain Reaction X Chromosome/genetics Y Chromosome/genetics N2 - Microchimerism has been implicated in the etiology of autoimmune diseases. It has also been implicated in the induction/maintenance of fetal tolerance. We used polymerase chain reaction (PCR) analysis to determine whether microchimerism occurred in patients who subsequently developed primary biliary cirrhosis (PBC), and thus may be involved in its etiology. We performed PCR amplification of sequences unique to both the X and Y chromosomes from the livers of 37 women with PBC and 39 female controls using WAVE technology; a very sensitive technology based on an ion-pair reverse-phase high-performance liquid chromatography system. All patients were known to have had at least 1 son and it was confirmed that PBC was diagnosed after the birth of the son. Data were analyzed for both detection of the Y chromosome gene and the ratio of the yield of the Y chromosome PCR products to the X chromosome. The prevalence of Y chromosome detection in PBC was 26 of 37 (70%) compared with 28 of 39 (72%) in controls, and the ratio of Y chromosome to X chromosome was similar between the PBC and control groups, 0.402 +/- 0.143 vs. 0.271 +/- 0.055, respectively. Our results, using our more sensitive technology, showed that microchimerism is a very common event in human liver and supported the thesis that this may contribute to the induction/maintenance of fetal tolerance. However, although we cannot exclude the possibility that select fetal major histocompatibility complex (MHC) haplotypes might contribute to disease susceptibility, our data suggest that microchimerism by itself does not play a significant role in the development of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10498630 ID - 128 ER - TY - JOUR AU - Reynoso-Paz, S. AU - Coppel, R. L. AU - Mackay, I. R. AU - Bass, N. M. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 1999 TI - The immunobiology of bile and biliary epithelium SP - 351-7 N1 - Aug JF - Hepatology JO - Hepatology (Baltimore, Md VL - 30 IS - 2 SN - 0270-9139 (Print) N1 - The immunobiology of bile and biliary epithelium N1 - 10421640 N1 - Dk39588/dk/niddk Gm17779/gm/nigms Journal Article Research Support, U.S. Gov't, P.H.S. Review United states N1 - eng KW - Animals Antigen-Presenting Cells/physiology Bile/*immunology Bile Ducts/*immunology Cell Communication Cytokines/physiology Epithelial Cells/immunology Humans Immunoglobulin A/analysis N2 - Long thought to be just a simple pipe involved in the delivery of bile from hepatocytes to the gallbladder and intestine, bile ducts are now regarded as highly dynamic structures consisting of cell populations involved in formation, transport and modification of bile by both secretory and absorptive processes. In fact, both bile and biliary epithelium appear to have active immunologic roles in both innate and adaptive immune responses. These roles are becoming increasingly clear as techniques have been developed allowing for the study of bile and biliary epithelial cells (BECs) in mucosal immunity. Bile is actively involved in the transport of immunoglobulin to the intestine, while BECs secrete chemokines and cytokines and serve to localize the immune response by expressing critical cell adhesion molecules. Evidence suggests that BECs may also function as professional antigen-presenting cells (APC) and, in the process, contribute to the modulation of inflammatory reactions. Bile ducts and, in particular, BECs, are the primary site of damage in several immunologically mediated liver diseases. Progress in these important areas has been rapid and forms the basis of this review. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Davis, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10421640 ID - 132 ER - TY - JOUR AU - Reynoso-Paz, S. AU - Coppel, R. L. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 1999 TI - Vanishing bile duct syndromes: considerations of the immunobiology of autoimmune biliary diseases SP - 37-44 N1 - Sep JF - Isr Med Assoc J VL - 1 IS - 1 SN - 1565-1088 (Print) N1 - Vanishing bile duct syndromes: considerations of the immunobiology of autoimmune biliary diseases N1 - 11370120 N1 - Journal Article Review Israel Imaj N1 - eng KW - Animals Autoimmune Diseases/*immunology Bile/chemistry/*immunology Bile Duct Diseases/*immunology Cell Adhesion Molecules Epithelial Cells/immunology Humans Immunoglobulin A, Secretory/metabolism Protein Transport AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Davis, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11370120 ID - 102 ER - TY - JOUR AU - Quaranta, S. AU - Van de Water, J. AU - Ishibashi, H. AU - Rosina, F. AU - Coppel, R. AU - Uibo, R. AU - Gershwin, M. E. PY - 1999 TI - The immunopathogenesis of primary biliary cirrhosis SP - 3041-7 N1 - Nov-Dec JF - Hepatogastroenterology JO - Hepato-gastroenterology VL - 46 IS - 30 SN - 0172-6390 (Print) N1 - The immunopathogenesis of primary biliary cirrhosis N1 - 10626157 N1 - Journal Article Review Greece N1 - eng KW - Autoantibodies/*immunology Autoantigens/*immunology Bile Ducts, Intrahepatic/immunology Epithelium/immunology Epitopes, T-Lymphocyte/immunology HLA Antigens/immunology Humans Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/immunology T-Lymphocytes/*immunology N2 - There have been several recent advances in our understanding of primary biliary cirrhosis. Foremost amongst these has been the cloning and identification of the mitochondrial autoantigens as members of the 2-oxo-dehydrogenase complex. These include the E2 components of PBC, BCKD and OGDC. The immunodominant autoepitopes of the autoantigens have been mapped and, in all cases, correspond to the inner lipoyl domain. Limited progress has also been made on T cells, particularly the CD4 response. However, the fundamental mechanisms and the role, if any, of CD8 cells are unknown. Finally, at least 2 groups have identified a molecule that cross-reacts with PDC-E2, i.e., a mimic, on the luminal surface of biliary epithelium in PBC but not controls. The identification of this molecule will be critical in further understanding the immune response of this disease. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, School of Medicine 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10626157 ID - 121 ER - TY - JOUR AU - Quaranta, S. AU - Shulman, H. AU - Ahmed, A. AU - Shoenfeld, Y. AU - Peter, J. AU - McDonald, G. B. AU - Van de Water, J. AU - Coppel, R. AU - Ostlund, C. AU - Worman, H. J. AU - Rizzetto, M. AU - Tsuneyama, K. AU - Nakanuma, Y. AU - Ansari, A. AU - Locatelli, F. AU - Paganin, S. AU - Rosina, F. AU - Manns, M. AU - Gershwin, M. E. PY - 1999 TI - Autoantibodies in human chronic graft-versus-host disease after hematopoietic cell transplantation SP - 106-16 N1 - Apr JF - Clin Immunol JO - Clinical immunology (Orlando, Fla VL - 91 IS - 1 SN - 1521-6616 (Print) N1 - Autoantibodies in human chronic graft-versus-host disease after hematopoietic cell transplantation N1 - 10219261 N1 - Ak 39588/phs Ca 15704/ca/nci Ca 18092/ca/nci Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Animals Antibodies, Anticardiolipin/blood Antibodies, Antinuclear/blood Antibody Specificity Autoantibodies/*blood Autoantigens Case-Control Studies Cattle Chronic Disease Female Graft vs Host Disease/etiology/*immunology Hematopoietic Stem Cell Transplantation/*adverse effects Humans Liver Cirrhosis, Biliary/etiology/immunology/pathology Liver Diseases/etiology/immunology/pathology Male Mitochondria/immunology Muscle, Smooth/immunology N2 - Primary biliary cirrhosis (PBC) and graft-versus-host disease (GVHD) are thought to have common immunopathologic features and previous studies have reported that 5.2 to 81% of patients with chronic GVHD after allogeneic hematopoietic cell transplant have antimitochondrial antibodies (AMA). We studied a total of 89 patients with chronic GVHD and 60 controls for AMA reactivity by ELISA and immunoblotting using recombinant PDC-E2, BCOADC-E2, and OGDC-E2, immunoblotting of beef heart mitochondrial proteins, and reactivity to nuclei, smooth muscle (ASMA), ribonucleoprotein JO-1, extractable nuclear antigen, nuclear proteins SSA/ SSB, ribonucleic P proteinase III, cardiolipin (ACA), liver kidney microsomal, thyroid microsomal, myeloperoxidase, and the reactivity of rheumatoid factor. A subset of 60 chronic GVHD sera were tested for reactivity to gp210 and LBR. Finally, liver tissue from patients with chronic GVHD and PBC was studied by immunohistochemistry to determine whether there was comparable abnormal apical staining of biliary epithelial cells using PDC-E2-specific monoclonal antibodies. Surprisingly, there were no AMA found in the sera from the 89 patients with chronic GVHD. Review of published data on AMA in GVHD suggests that previous results were primarily false positives. In contrast, sera from the patients with GVHD did have a variety of other autoantibodies and, in particular, 20/89 (22.4%) positive ANA, 23/89 (25.8%) positive ASMA, and 9/89 (10.1%) positive ACA. The other autoantibodies assayed were not statistically different from controls. Finally, abnormal biliary epithelial luminal staining of bile ducts was found, as expected, in liver tissue of patients with PBC but was absent in chronic GVHD. AD - University of California, Davis 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10219261 ID - 137 ER - TY - JOUR AU - Nalbandian, G. AU - Van de Water, J. AU - Gish, R. AU - Manns, M. AU - Coppel, R. L. AU - Rudich, S. M. AU - Prindiville, T. AU - Gershwin, M. E. PY - 1999 TI - Is there a serological difference between men and women with primary biliary cirrhosis? SP - 2482-6 N1 - Sep JF - Am J Gastroenterol JO - The American journal of gastroenterology VL - 94 IS - 9 SN - 0002-9270 (Print) N1 - Is there a serological difference between men and women with primary biliary cirrhosis? N1 - 10484012 N1 - Dk 39588/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Female Humans Liver Cirrhosis, Biliary/*blood Male Middle Aged Sex Characteristics Sex Factors N2 - OBJECTIVE: Primary biliary cirrhosis (PBC) is an autoimmune disease affecting small intrahepatic bile ducts of the liver, causing destruction of the epithelium that results in eventual fibrosis and scarring. We still lack a complete epidemiological description of this disease, although interesting geographic differences in prevalence have been described. One consistent feature has been the relative scarcity of men with PBC. In fact, published ratios of women to men range from 3:1 to as high as 22:1. Thus far, the only clinical difference reported between men and women with PBC is a putative higher risk of hepatocarcinoma in men. Previous serological studies have shown that about 95% of all patients possess antimitochondrial antibodies to members of the highly conserved 2-oxo-acid dehydrogenase family of proteins, namely pyruvate dehydrogenase complex E2 (PDC-E2), branched-chain 2-oxo-acid dehydrogenase complex E2 (BCOADC-E2), and 2-oxo glutarate dehydrogenase complex E2 (OGDC-E2). However, there has been no information as to whether there is a difference in serological response between men and women. Using the serological hallmark of antimitochondrial antibodies (AMAs) and taking advantage of the availability of recombinant mitochondrial autoantigens, investigations were performed to determine if there were any serological differences between men and women with PBC. METHODS: Sera were collected from 88 patients with PBC, of whom 46 were men and 42 were women. Using a combination of immunoblotting and enzyme-linked immunoabsorbent assay (ELISA) against beef heart mitochondria (BHM), recombinant PDC-E2, BCOADC-E2, and OGDC-E2, we determined the relative autoantibody reactivities of our study population. RESULTS: Both men and women with PBC produced high titer antimitochondrial antibodies. The frequency of reactivity was similar in both groups and included, in descending order, PDC-E2, E3BP (Protein X), BCOADC-E2, and finally OGDC-E2. More importantly, antigenic specificity was nearly identical regardless of gender. CONCLUSIONS: AMAs are the serological hallmark of PBC in both men and women, and there is no significant difference in reactivity between the two groups of patients. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California at Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10484012 ID - 129 ER - TY - JOUR AU - Kun, J. F. AU - Waller, K. L. AU - Coppel, R. L. PY - 1999 TI - Plasmodium falciparum: structural and functional domains of the mature-parasite-infected erythrocyte surface antigen SP - 258-67 N1 - Mar JF - Exp Parasitol JO - Experimental parasitology VL - 91 IS - 3 SN - 0014-4894 (Print) N1 - Plasmodium falciparum: structural and functional domains of the mature-parasite-infected erythrocyte surface antigen N1 - 10072328 N1 - Dk 32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*chemistry/genetics/metabolism Cytoskeleton/metabolism DNA, Complementary/chemistry DNA, Protozoan/chemistry Exons Molecular Sequence Data Plasmodium falciparum/genetics/*immunology Point Mutation Polymerase Chain Reaction Protozoan Proteins/*chemistry/genetics/metabolism RNA, Messenger/genetics Sequence Deletion N2 - The mature parasite-infected erythrocyte surface antigen (MESA) is a protein exported to the membrane skeleton of the infected red cell, where it forms a strong noncovalent interaction with the host red cell protein, protein 4.1. The complete gene structure of MESA from the Ugandan isolate Palo Alto is described. Comparison to the previously reported MESA sequence from the Papua New Guinean cloned line D10 reveals strong conservation of the general gene structure of a short first exon and a long second exon. The exact exon/intron boundaries were determined by the generation and sequencing of a cDNA from this region. The MESA gene from both isolates consists of seven blocks of repeats that are identical in order. Repeat blocks are conserved to a high degree; however, differences are noted in most blocks in the form of scattered mutations or differences in repeat numbers. Previous work had shown that synthetic peptides spanning a 19-residue region could inhibit the binding of MESA to protein 4.1. Removal of this region from MESA almost completely abolished the binding of MESA to IOVs. Sequencing of this region from a number of laboratory and field isolates demonstrates complete conservation of the cytoskeletal binding domain and flanking sequences. AD - Institut fur Tropenmedizin, Wilhelmstrasse 27, Tubingen, 72074, Germany. juergen.kun@uni-tuebingen.de UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10072328 ID - 140 ER - TY - JOUR AU - Katayanagi, K. AU - Van de Water, J. AU - Kenny, T. AU - Nakanuma, Y. AU - Ansari, A. A. AU - Coppel, R. AU - Gershwin, M. E. PY - 1999 TI - Generation of monoclonal antibodies to murine bile duct epithelial cells: identification of annexin V as a new marker of small intrahepatic bile ducts SP - 1019-25 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 29 IS - 4 SN - 0270-9139 (Print) N1 - Generation of monoclonal antibodies to murine bile duct epithelial cells: identification of annexin V as a new marker of small intrahepatic bile ducts N1 - 10094941 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Amino Acid Sequence Animals Annexin A5/immunology/*metabolism Antibodies, Monoclonal/*biosynthesis Antibody Specificity Bile Ducts, Intrahepatic/*metabolism Biological Markers/analysis Cells, Cultured Epithelial Cells/immunology/metabolism Female Hybridomas/immunology/metabolism Immunohistochemistry Mice Mice, Inbred BALB C Molecular Sequence Data Molecular Weight Organ Specificity Precipitin Tests Rats Sequence Analysis N2 - Biliary epithelial cells (BECs) are distributed along the length of both the extrahepatic and intrahepatic biliary tree, but have distinctly different phenotypes and functions according to their anatomical location. It has been reasoned that the distinct appearance of pathogenic lesions in different biliary diseases may be associated with the expression of distinct proteins. These data prompted us to immunize rats with cultured murine BECs with the objective of determining if there are unique antigens on BECs. Of the 45 monoclonal antibodies (mAbs) produced, 12 mAbs (MBEC 1-12) were selected for detailed study based on their classification into three major groups. These groups included four antibodies (MBEC 1-4) that reacted in a staining pattern typical of mucin. A second group of mAbs, MBECs 5 to 8, reacted strongly along the biliary tract and by immunoblot analysis, reacted with several bands ranging from 44 kd to 64 kd. These antibodies were considered as markers of pan BECs and their staining pattern proved similar to that of a control polyclonal pan-cytokeratin. The final group of mAbs, MBECs 9 to 12, recognized a 36-kd antigen using lysates of murine BECs. These antibodies also predominantly stained small peripheral bile ducts. The reactive antigen was purified by immunoprecipitation and microsequenced; the peptides sequenced showed 100% homology with murine annexin V. The identification of annexin V with predominantly intrahepatic bile ducts, is of significant interest because of the multiple roles of annexin V, including that of membrane cytoskeletal interactions during transport and apoptosis. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10094941 ID - 138 ER - TY - JOUR AU - Eisen, D. P. AU - Marshall, V. M. AU - Billman-Jacobe, H. AU - Coppel, R. L. PY - 1999 TI - A Plasmodium falciparum apical membrane antigen-1 (AMA-1) gene apparently generated by intragenic recombination SP - 243-6 N1 - May 25 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 100 IS - 2 SN - 0166-6851 (Print) N1 - A Plasmodium falciparum apical membrane antigen-1 (AMA-1) gene apparently generated by intragenic recombination N1 - 10391387 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Amino Acid Substitution Animals *Antigens, Protozoan Genes, Protozoan Membrane Proteins/chemistry/*genetics Molecular Sequence Data Plasmodium falciparum/chemistry/*genetics Protozoan Proteins/chemistry/*genetics *Recombination, Genetic AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10391387 ID - 133 ER - TY - JOUR AU - Dubel, L. AU - Tanaka, A. AU - Leung, P. S. AU - Van de Water, J. AU - Coppel, R. AU - Roche, T. AU - Johanet, C. AU - Motokawa, Y. AU - Ansari, A. AU - Gershwin, M. E. PY - 1999 TI - Autoepitope mapping and reactivity of autoantibodies to the dihydrolipoamide dehydrogenase-binding protein (E3BP) and the glycine cleavage proteins in primary biliary cirrhosis SP - 1013-8 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 29 IS - 4 SN - 0270-9139 (Print) N1 - Autoepitope mapping and reactivity of autoantibodies to the dihydrolipoamide dehydrogenase-binding protein (E3BP) and the glycine cleavage proteins in primary biliary cirrhosis N1 - 10094940 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Amino Acid Oxidoreductases/*immunology Amino Acid Sequence Animals Antibody Specificity Autoantibodies/*immunology Carrier Proteins/*immunology Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay *Epitope Mapping Humans Immunoblotting Liver Cirrhosis, Biliary/enzymology/*immunology Mitochondria/immunology Molecular Sequence Data Multienzyme Complexes/*immunology Peptides/*immunology Pyruvate Dehydrogenase Complex/*immunology/metabolism Recombinant Proteins/immunology/metabolism Thioctic Acid/biosynthesis Transferases/*immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the presence of antimitochondrial antibodies (AMA) directed primarily against the E2 subunits of the pyruvate dehydrogenase complex, the branched chain 2-oxo-acid dehydrogenase complex, the 2-oxoglutarate dehydrogenase complex, as well as the dihydrolipoamide dehydrogenase-binding protein (E3BP) of pyruvate dehydrogenase complex. The autoantibody response to each E2 subunit is directed to the lipoic acid binding domain. However, hitherto, the epitope recognized by autoantibodies to E3BP has not been mapped. In this study, we have taken advantage of the recently available full-length human E3BP complementary DNA (cDNA) to map this epitope. In addition, another lipoic binding protein, the H-protein of the glycine cleavage complex, was also studied as a potential autoantigen recognized by AMA. Firstly, the sequence corresponding to the lipoic domain of E3BP (E3BP-LD) was amplified by polymerase chain reaction and recombinant protein and then purified. Immunoreactivity of 45 PBC sera (and 52 control sera) against the purified recombinant E3BP-LD was analyzed by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Secondly, reactivity of PBC sera was similarly analyzed by immunoblotting against H-protein. It is interesting that preabsorption of patient sera with the lipoic acid binding domain of E3BP completely removed all reactivity with the entire protein by immunoblotting analysis, suggesting that autoantibodies to E3BP are directed solely to its lipoic acid binding domain. Fifty-three percent of PBC sera reacted with E3BP-LD, with the majority of the response being of the immunoglobulin G (IgG) isotype (95%). Surprisingly, there was little IgM response to the E3BP-LD suggesting that the immune response was secondary because of determinant spreading. In contrast, H-protein does not appear to possess (or expose) autoepitopes recognized by PBC sera. This observation is consistent with structural data on this moiety. AD - Department of Microbiology, Monash University, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10094940 ID - 139 ER - TY - JOUR AU - Black, C. G. AU - Wang, L. AU - Hibbs, A. R. AU - Werner, E. AU - Coppel, R. L. PY - 1999 TI - Identification of the Plasmodium chabaudi homologue of merozoite surface proteins 4 and 5 of Plasmodium falciparum SP - 2075-81 N1 - May JF - Infect Immun JO - Infection and immunity VL - 67 IS - 5 SN - 0019-9567 (Print) N1 - Identification of the Plasmodium chabaudi homologue of merozoite surface proteins 4 and 5 of Plasmodium falciparum N1 - 10225857 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence DNA Primers/genetics DNA, Protozoan/genetics Erythrocytes/parasitology Female Gene Expression Genes, Protozoan Membrane Proteins/*genetics Mice Mice, Inbred BALB C Molecular Sequence Data Plasmodium chabaudi/*genetics Plasmodium falciparum/*genetics Protozoan Proteins/*genetics Sequence Homology, Amino Acid Species Specificity N2 - Previous studies of Plasmodium falciparum have identified a region of chromosome 2 in which are clustered three genes for glycosylphosphatidylinositol (GPI)-anchored merozoite surface proteins, MSP2, MSP5, and MSP4, arranged in tandem. MSP4 and MSP5 both encode proteins 272 residues long that contain hydrophobic signal sequences, GPI attachment signals, and a single epidermal growth factor (EGF)-like domain at their carboxyl termini. Nevertheless, the remainder of their protein coding regions are quite dissimilar. The locations and similar structural features of these genes suggest that they have arisen from a gene duplication event. Here we describe the identification of the syntenic region of the genome in the murine malaria parasite, Plasmodium chabaudi adami DS. Only one open reading frame is present in this region, and it encodes a protein with structural features reminiscent of both MSP4 and MSP5, including a single EGF-like domain. Accordingly, the gene has been designated PcMSP4/5. The homologue of the P. falciparum MSP2 gene could not be found in P. chabaudi; however, the amino terminus of the PcMSP4/5 protein shows similarity to that of MSP2. The PcMSP4/5 gene encodes a protein with an apparent molecular mass of 36 kDa, and this protein is detected in mature stages of the parasite. The protein partitions in the detergent-enriched phase after Triton X-114 fractionation and is localized to the surfaces of trophozoites and developing and free merozoites. The PcMSP4/5 gene is transcribed in both ring and trophozoite stages but appears to be spliced in a stage-specific manner such that the central intron is spliced from the mRNA in the parasitic stage in which the protein is expressed. AD - Department of Microbiology, Monash University, Clayton 3168, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10225857 ID - 136 ER - TY - JOUR AU - Billman-Jacobe, H. AU - McConville, M. J. AU - Haites, R. E. AU - Kovacevic, S. AU - Coppel, R. L. PY - 1999 TI - Identification of a peptide synthetase involved in the biosynthesis of glycopeptidolipids of Mycobacterium smegmatis SP - 1244-53 N1 - Sep JF - Mol Microbiol JO - Molecular microbiology VL - 33 IS - 6 SN - 0950-382X (Print) N1 - Identification of a peptide synthetase involved in the biosynthesis of glycopeptidolipids of Mycobacterium smegmatis N1 - 10510238 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Alleles Amino Acid Sequence Base Sequence DNA Primers/genetics Genes, Bacterial Glycoconjugates/*biosynthesis/chemistry Molecular Sequence Data Mutagenesis, Insertional Mutation Mycobacterium smegmatis/enzymology/genetics/*metabolism Peptide Synthases/genetics/*metabolism Sequence Homology, Amino Acid N2 - Five rough colony mutants of Mycobacterium smegmatis mc2155 were produced by transposon mutagenesis. The mutants were unable to synthesize glycopeptidolipids that are normally abundant in the cell wall of wild-type M. smegmatis. The glycopeptidolipids have a lipopeptide core comprising a fatty acid amide linked to a tetrapeptide that is modified with O-methylated rhamnose and O-acylated 6-deoxy talose. Compositional analysis of lipids extracted from the mutants indicated that the defect in glycopeptidolipid synthesis occurred in the assembly of the lipopeptide core. No other defects or compensatory changes in cell wall structure were detected in the mutants. All five mutants had transposon insertions in a gene encoding an enzyme belonging to the peptide synthetase family. Targeted disruption of the gene in the wild-type strain gave a phenotype identical to that of the five transposon mutants. The M. smegmatis peptide synthetase gene is predicted to encode four modules that each contain domains for cofactor binding and for amino acid recognition and adenylation. Three modules also have amino acid racemase domains. These data suggest that the common lipopeptide core of these important cell wall glycolipids is synthesized by a peptide synthetase. AD - Department of Microbiology, Monash University, Clayton, Victoria, 3168, Australia. Helen.BJ@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10510238 ID - 127 ER - TY - JOUR AU - Billman-Jacobe, H. AU - Haites, R. E. AU - Coppel, R. L. PY - 1999 TI - Characterization of a Mycobacterium smegmatis mutant lacking penicillin binding protein 1 SP - 3011-3 N1 - Dec JF - Antimicrob Agents Chemother JO - Antimicrobial agents and chemotherapy VL - 43 IS - 12 SN - 0066-4804 (Print) N1 - Characterization of a Mycobacterium smegmatis mutant lacking penicillin binding protein 1 N1 - 10582900 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - *Bacterial Proteins Carrier Proteins/*genetics Cell Membrane Permeability/genetics Culture Media Glycine/metabolism Hexosyltransferases/genetics/metabolism Kinetics Multienzyme Complexes/genetics/metabolism Muramoylpentapeptide Carboxypeptidase/*genetics Mutation/*genetics Mycobacterium smegmatis/enzymology/*genetics/growth & development Penicillin-Binding Proteins Peptidyl Transferases/genetics/metabolism Polymerase Chain Reaction beta-Lactamases/genetics/metabolism N2 - The ponA gene of Mycobacterium smegmatis encodes a 95-kDa penicillin binding protein, PBP1, that is similar to PBP1s of Mycobacterium tuberculosis and Mycobacterium leprae. Transposon disruption of ponA in M. smegmatis resulted in a PBP1-deficient mutant that was sensitive to beta-lactam antibiotics, was more permeable to glycine, and grew slowly in liquid culture. AD - Department of Microbiology, Monash University, Clayton, Victoria 3168, Australia. Helen.BJ@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10582900 ID - 124 ER - TY - JOUR AU - Yeung, J. AU - Brown, J. T. AU - Nair, A. AU - Meites, E. AU - Coppel, R. L. AU - Mohandas, N. AU - Meyer-Ilse, W. AU - Magowan, C. PY - 1998 TI - X-ray microscopic visualization of specific labeling of adhesive molecule CD36 and cytoadherence by Plasmodium falciparum infected erythrocytes SP - 245-58 N1 - Mar JF - Res Commun Mol Pathol Pharmacol JO - Research communications in molecular pathology and pharmacology VL - 99 IS - 3 SN - 1078-0297 (Print) N1 - X-ray microscopic visualization of specific labeling of adhesive molecule CD36 and cytoadherence by Plasmodium falciparum infected erythrocytes N1 - 9591321 N1 - Dk32094-10/dk/niddk Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Antigens, CD36/*analysis/immunology Cell Adhesion Electron Probe Microanalysis Endothelium, Vascular/immunology/parasitology Erythrocytes/*parasitology/physiology/ultrastructure Fluorescent Antibody Technique, Indirect Humans Melanoma/immunology/parasitology/ultrastructure Plasmodium falciparum/*physiology/ultrastructure Tumor Cells, Cultured N2 - We investigated the cytoadherence of Plasmodium falciparum infected erythrocytes to target cells that express CD36 by soft x-ray microscopy. Using immunogold beads enhanced with silver, we localized CD36 on the surface of intact melanoma cells and throughout Triton extracted melanoma cells. We examined the orientation of parasites within erythrocytes that bound to target cells, and the interactions between the red cell membrane and the target cell, and we confirmed that fibrillar structures on the surface of melanoma and endothelial cells can be involved in the association between infected erythrocytes and melanoma cells or endothelial cells. AD - Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9591321 ID - 149 ER - TY - JOUR AU - Shimoda, S. AU - Van de Water, J. AU - Ansari, A. AU - Nakamura, M. AU - Ishibashi, H. AU - Coppel, R. L. AU - Lake, J. AU - Keeffe, E. B. AU - Roche, T. E. AU - Gershwin, M. E. PY - 1998 TI - Identification and precursor frequency analysis of a common T cell epitope motif in mitochondrial autoantigens in primary biliary cirrhosis SP - 1831-40 N1 - Nov 15 JF - J Clin Invest JO - The Journal of clinical investigation VL - 102 IS - 10 SN - 0021-9738 (Print) N1 - Identification and precursor frequency analysis of a common T cell epitope motif in mitochondrial autoantigens in primary biliary cirrhosis N1 - 9819369 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged Autoantigens/*immunology Cell Division Cross Reactions Dihydrolipoyllysine-Residue Acetyltransferase Epitopes, T-Lymphocyte/*immunology Female Humans Leukocytes, Mononuclear/cytology Liver/immunology Liver Cirrhosis, Biliary/*immunology Lymph Nodes/immunology Middle Aged Mitochondria/*immunology Peptides/immunology Pyruvate Dehydrogenase Complex/*immunology T-Lymphocytes/immunology N2 - The immunodominant antimitochondrial antibody response in patients with primary biliary cirrhosis (PBC) is directed against the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Based on our earlier observations regarding peripheral blood mononuclear cell (PBMC) T cell epitopes, we reasoned that a comparative analysis of the precursor frequencies of PDC-E2 163-176-specific T cells isolated from PBMC, regional hepatic lymph nodes, and from the liver of PBC patients would provide insight regarding the role of T cells in PBC. Results showed a disease-specific 100-150-fold increase in the precursor frequency of PDC-E2 163-176-specific T cells in the hilar lymph nodes and liver when compared with PBMC from PBC patients. Interestingly, autoreactive T cells and autoantibodies from PBC patients both recognize the same dominant epitope. In addition, we demonstrated cross-reactivity of PDC-E2 peptide 163-176-specific T cell clones with PDC-E2 peptide 36-49 and OGDC-E2 peptide 100-113 thereby identifying a common T cell epitope "motif" ExETDK. The peptide 163-176-specific T cell clones also reacted with purified native PDC-E2, suggesting that this epitope is not a cryptic determinant. These data provide evidence for a major role for PDC-E2 peptide 163-176 and/or peptides bearing a similar motif in the pathogenesis of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California, Davis, California 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9819369 ID - 143 ER - TY - JOUR AU - Migliaccio, C. AU - Nishio, A. AU - Van de Water, J. AU - Ansari, A. A. AU - Leung, P. S. AU - Nakanuma, Y. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1998 TI - Monoclonal antibodies to mitochondrial E2 components define autoepitopes in primary biliary cirrhosis SP - 5157-63 N1 - Nov 15 JF - J Immunol VL - 161 IS - 10 SN - 0022-1767 (Print) N1 - Monoclonal antibodies to mitochondrial E2 components define autoepitopes in primary biliary cirrhosis N1 - 9820485 N1 - Dk39558/dk/niddk Dk50977/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - Animals Antibodies, Monoclonal/*chemistry Antibody Specificity Antigen-Antibody Reactions Autoantibodies/*chemistry Autoantigens/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Epitopes/immunology Female Humans Immunohistochemistry Liver Cirrhosis, Biliary/enzymology/*immunology Mice Mice, Inbred BALB C Mitochondria, Liver/enzymology/*immunology Pyruvate Dehydrogenase Complex/*immunology Tumor Cells, Cultured N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the presence of antimitochondrial Abs (AMA). The autoantigens recognized by AMA are the E2 components of the pyruvate dehydrogenase complex (PDC-E2), the branched chain 2-oxoacid dehydrogenase complex E (BCOADC-E2), and the 2-oxoglutarate dehydrogenase complex E (OGDC-E2). Previous studies using murine monoclonal and human combinatorial Abs to PDC-E2 have demonstrated an intense linear staining pattern in the apical region of biliary epithelial cells (BEC) in PBC but not control liver. We therefore examined whether mAbs to the other mitochondrial autoantigens BCOADC-E2 and OGDC-E2 demonstrated disease-specific patterns of reactivity. Using an expressed recombinant "trihybrid" protein containing the lipoyl domains of PDC-E2, OGDC-E2, and BCOADC-E2, we immunized BALB/c mice to produce 35 mAbs specific for one or more of the above mitochondrial autoantigens. Seven of these mAbs uniquely stained the apical region of BEC in PBC. Of these seven, one was reactive to PDC-E2, two recognized BCOADC-E2, three were reactive to OGDC-E2, and one recognized all three Ags. Our current data demonstrate that, similar to our previous studies regarding PDC-E2, mAbs to BCOADC-E2 and OGDC-E2, or a molecule that cross-reacts with the inner lipoyl domain of all three enzymes, also show a uniquely intense staining pattern in the apical region of BEC in patients with PBC when compared with diseased controls. The abundance of such disease-specific determinants in the target cells of PBC raises interesting possibilities regarding the role of these autoantigens in the pathogenesis of this disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, Davis 95616-8660, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9820485 ID - 142 ER - TY - JOUR AU - Mattalia, A. AU - Quaranta, S. AU - Leung, P. S. AU - Bauducci, M. AU - Van de Water, J. AU - Calvo, P. L. AU - Danielle, F. AU - Rizzetto, M. AU - Ansari, A. AU - Coppel, R. L. AU - Rosina, F. AU - Gershwin, M. E. PY - 1998 TI - Characterization of antimitochondrial antibodies in health adults SP - 656-61 N1 - Mar JF - Hepatology JO - Hepatology (Baltimore, Md VL - 27 IS - 3 SN - 0270-9139 (Print) N1 - Characterization of antimitochondrial antibodies in health adults N1 - 9500690 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged Autoantibodies/*blood Epitope Mapping Female Humans Immunoglobulin G/classification Immunoglobulin Isotypes/blood Liver Cirrhosis, Biliary/*diagnosis Male Middle Aged Mitochondria/*immunology Recombinant Proteins/immunology N2 - The detection of antimitochondrial antibodies (AMAs) is an important criterion for the diagnosis of primary biliary cirrhosis (PBC). During the last decade, the mitochondrial autoantigens have been cloned, sequenced, and identified as members of the 2-oxo-acid dehydrogenase pathway, including the E2 subunits of pyruvate dehydrogenase (PDC-E2), branched-chain 2-oxo-acid dehydrogenase (BCOADC-E2), and 2-oxo-glutarate dehydrogenase (OGDC-E2). We have developed a rapid and sensitive diagnostic test for use in PBC based on a triple hybrid recombinant molecule (r-MIT3) that contains the autoepitopes of PDC-E2, BCOADC-E2, and OGDC-E2. To help understand the frequency and antigen specificity of AMAs in an asymptomatic population and to identify patients with early disease, we investigated the prevalence of AMA, by enzyme-linked immunosorbent assay (ELISA), in a cohort of 1,530 people from northern Italy. Positive sera were further analyzed for immunoglobulin (Ig) isotypes, subclasses, and epitopes of AMA by a combination of ELISA and immunoblotting. In this cohort of 1,530 people, 9 (0.5%) reacted to r-MIT3 by ELISA. Of the 9 reactive sera, 2 recognized PDC-E2, 2 of 9 recognized BCOADC-E2, 1 of 9 recognized OGDC-E2, 2 of 9 recognized both PDC-E2 and BCOADC-E2, and 1 of 9 recognized PDC-E2 and OGDC-E2. AMA reactivity was primarily IgM and IgA. Epitope mapping revealed an AMA pattern of reactivity to PDC-E2 that differed from that found in patients with histologically proven PBC in most of the sera. However, 1 sera of a 72-year-old female with a normal alkaline phosphatase had an AMA profile identical to typical PBC. After a variable follow-up period (8-14 months), sera from 8 of 9 of these people were re-obtained for AMA and relative epitope mapping. Interestingly, the reactivity had a wider AMA pattern than before. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9500690 ID - 151 ER - TY - JOUR AU - Masanaga, T. AU - Watanabe, Y. AU - Van de Water, J. AU - Leung, P. S. AU - Nakanishi, T. AU - Kajiyama, G. AU - Ruebner, B. H. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1998 TI - Induction and persistence of immune-mediated cholangiohepatitis in neonatally thymectomized mice SP - 141-9 N1 - Nov JF - Clin Immunol Immunopathol JO - Clinical immunology and immunopathology VL - 89 IS - 2 SN - 0090-1229 (Print) N1 - Induction and persistence of immune-mediated cholangiohepatitis in neonatally thymectomized mice N1 - 9787116 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adoptive Transfer Animals *Animals, Newborn Antibodies/analysis Autoimmune Diseases/etiology Cholangitis/*immunology/pathology Female Fluorescent Antibody Technique Hepatitis, Animal/*immunology Immunoblotting Liver/pathology Mice Mice, Inbred A Mitochondria, Liver/immunology *Thymectomy N2 - The availability of recombinant autoantigens allows the experimental study of the relationships between primary biliary cirrhosis (PBC) and mitochondrial antigens. We took advantage of these recombinant autoantigens and attempted to induce autoimmune cholangitis by immunizing neonatally thymectomized (NTx) lipopolysaccharide (LPS)-treated A/J mice, known to be prone to organ-specific autoimmune diseases. We employed a recombinant protein containing a dual-headed molecule that coexpresses the immunodominant epitope of the E2 subunits of the pyruvate dehydrogenase complex and the branched-chain keto-acid dehydrogenase complex. We report herein that an immune-mediated cholangiohepatitis was induced by such immunization and the concurrent injection of LPS into NTx mice. The incidence of cholangitis was 79% in the NTx, immunized, LPS group compared to 14% in the NTx, nonimmunized, LPS group. The histopathology ranged from mild to severe and included bile duct damage, focal hepatic necrosis, and endotheliitis, but no granulomas. Moreover, almost all such lesions persisted for 12 weeks after the discontinuation of immunization and LPS injections in the NTx mice. Interestingly, we were successful (89%) in transferring the cholangiohepatitis by injection of liver infiltrating mononuclear cells from the NTx, immunized, LPS mice into congenic nonimmunized NTx mice; such lesions could not be transferred with spleen cells. Although the pathology is not typical of PBC, this model offers a unique venue for the study of immune-mediated hepatobiliary injury. AD - First Department of Internal Medicine, Hiroshima University School of Medicine, Hiroshima, 730, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9787116 ID - 145 ER - TY - JOUR AU - Marshall, V. M. AU - Tieqiao, W. AU - Coppel, R. L. PY - 1998 TI - Close linkage of three merozoite surface protein genes on chromosome 2 of Plasmodium falciparum SP - 13-25 N1 - Jul 1 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 94 IS - 1 SN - 0166-6851 (Print) N1 - Close linkage of three merozoite surface protein genes on chromosome 2 of Plasmodium falciparum N1 - 9719507 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan *Antigens, Protozoan Antimalarials Base Sequence Chromosomes/*genetics DNA, Complementary DNA, Protozoan Drug Combinations Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique, Indirect *Genes, Protozoan Immunoblotting Mefloquine/analogs & derivatives Membrane Proteins/*genetics Molecular Sequence Data Plasmodium falciparum/*genetics/isolation & purification Polymerase Chain Reaction Protozoan Proteins/*genetics Pyrimethamine Rabbits Sulfadoxine N2 - We have analysed a 10.5 kb region of chromosome 2 in Plasmodium falciparum that encompasses the coding region of four genes. Three genes are arranged in a head-to-tail orientation and encode the merozoite surface proteins MSP2 and MSP4 as well as a previously unreported sequence that encodes a polypeptide with the characteristics of a merozoite surface protein, now designated MSP5. The fourth gene, asl, is arranged in a tail-to-tail orientation with msp2 and has homology with prokaryotic and eukaryotic genes encoding adenylosuccinate lyase (ASL), an enzyme involved in purine biosynthesis and salvage. The genes, arranged in the order msp4, msp5, msp2 and asl, are separated by intergenic distances of 1021, 1017 and 722 bp, respectively. msp4 and msp5 are clearly related genes, each being composed of 2 exons and encoding proteins of identical length. Both msp4 and msp5 encode proteins that contain hydrophobic signal sequences, apparent glycosylphosphatidylinositol (GPI) attachment signals and a single epidermal growth factor-like (EGF-like) domain at their carboxyl termini. Nevertheless, the remainder of their protein coding regions are quite dissimilar. It appears that one of these genes arose as a result of a relatively ancient gene duplication event and both genes have subsequently diverged considerably. This study shows that msp5 is transcribed in asexual stages and its encoded product is a 40 kDa protein that appears to be located on the merozoite surface as determined by immunofluorescence assays. AD - The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. vmarshal@wehi.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9719507 ID - 147 ER - TY - JOUR AU - Malmborg, A. C. AU - Shultz, D. B. AU - Luton, F. AU - Mostov, K. E. AU - Richly, E. AU - Leung, P. S. AU - Benson, G. D. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. AU - Van de Water, J. PY - 1998 TI - Penetration and co-localization in MDCK cell mitochondria of IgA derived from patients with primary biliary cirrhosis SP - 573-80 N1 - Oct JF - J Autoimmun JO - Journal of autoimmunity VL - 11 IS - 5 SN - 0896-8411 (Print) N1 - Penetration and co-localization in MDCK cell mitochondria of IgA derived from patients with primary biliary cirrhosis N1 - 9802945 N1 - Ai 25144/ai/niaid Ai 31585/ai/niaid Dk39588/dk/niddk In Vitro Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Autoantigens Autoimmune Diseases/enzymology/*immunology Biological Transport, Active Case-Control Studies Cell Line Dihydrolipoyllysine-Residue Acetyltransferase Dogs Humans Immunoglobulin A/*metabolism Liver/immunology Liver Cirrhosis, Biliary/enzymology/*immunology Mice Microscopy, Fluorescence Mitochondria/enzymology/*immunology Pyruvate Dehydrogenase Complex/immunology/metabolism Receptors, Fc/genetics/metabolism Transfection N2 - Primary biliary cirrhosis (PBC) is a chronic autoimmune liver disease of unknown etiology characterized by high-titer anti-mitochondrial antibodies. The major autoantigen has been identified as the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2). The fact that PDC-E2 is present in all nucleated cells, but autoimmune damage is confined to biliary epithelial cells, prompted us to investigate the possibility that mucosally-derived IgA may be pathogenic for biliary epithelial cells. Serum IgA was purified from six patients with PBC and its localization and ability to penetrate cells was studied using Madine-Darby canine kidney (MDCK) cells transfected with the human IgA receptor (MDCK-pIgR). The potential of IgA to be transported through the cells was studied by a combination of immunohistochemistry and dual color fluorescent microscopy. Interestingly, IgA from all PBC patients co-localized with PDC-E2 (the major autoantigen of PBC) inside the cells; this was demonstrated by dual staining with anti-human IgA and a mouse monoclonal antibody directed to PDC-E2. In contrast, no co-localization was observed for IgA controls. Furthermore, dual staining of liver sections from PBC patients demonstrated co-localization of IgA and PDC-E2, both cytoplasmically and at the apical surface. We postulate that there may be a direct effect of these autoantibodies on the mitochondrial function of biliary epithelial cells. AD - Division of Rheumatology/Allergy Clinical Immunology, University of California, Davis, California, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9802945 ID - 144 ER - TY - JOUR AU - Magowan, C. AU - Liang, J. AU - Yeung, J. AU - Takakuwa, Y. AU - Coppel, R. L. AU - Mohandas, N. PY - 1998 TI - Plasmodium falciparum: influence of malarial and host erythrocyte skeletal protein interactions on phosphorylation in infected erythrocytes SP - 40-9 N1 - May JF - Exp Parasitol JO - Experimental parasitology VL - 89 IS - 1 SN - 0014-4894 (Print) N1 - Plasmodium falciparum: influence of malarial and host erythrocyte skeletal protein interactions on phosphorylation in infected erythrocytes N1 - 9603487 N1 - Dk 32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Blotting, Western *Cytoskeletal Proteins Detergents Erythrocyte Membrane/metabolism Erythrocytes/metabolism/*parasitology Humans Membrane Proteins/chemistry/*metabolism Molecular Weight *Neuropeptides Octoxynol Phenotype Phosphorylation Phosphotransferases/analysis Plasmodium falciparum/*metabolism Precipitin Tests Protozoan Proteins/chemistry/*metabolism Solubility N2 - Phosphorylation of components of the erythrocyte membrane skeleton has major effects on the physical properties of the membrane. Infection of red cells by the protozoan parasite Plasmodium falciparum leads to a marked increase in the level of phosphorylation of red cell protein 4.1 and the insertion into the red cell skeleton of parasite-encoded phosphoproteins, including the mature-parasite-infected erythrocyte surface antigen (MESA). Because of the tight association of MESA with protein 4.1, we set out to determine the importance of this interaction and that of other parasite-encoded skeletal-associated proteins to phosphorylation of the infected red cell membrane. Our results show that neither MESA nor protein 4.1 is required for phosphorylation of its binding partner. Further, phosphorylation of MESA and protein 4.1 occurs independently of the presence of knobs, the expression of PfHRP1, or cytoadherence phenotype. In contrast to previous studies, we were unable to detect a change in the molecular weight of protein 4.1 in erythrocytes infected with cytoadherent parasite lines. In red cells infected with parasites expressing PfHRP1 (K+), MESA and protein 4.1 are substrates for a kinase with the inhibitor profile of a casein kinase. Surprisingly, however, when we examined phosphorylation of MESA and protein 4.1 in K(-)-infected erythrocytes, we found that casein kinase I and II inhibitors had no, or greatly reduced, effectiveness, and in fact, phosphorylation of these two proteins was enhanced in some instances. AD - Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA. cathie_magowan@macmail2.lbl.gov UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9603487 ID - 148 ER - TY - JOUR AU - Eisen, D. AU - Billman-Jacobe, H. AU - Marshall, V. F. AU - Fryauff, D. AU - Coppel, R. L. PY - 1998 TI - Temporal variation of the merozoite surface protein-2 gene of Plasmodium falciparum SP - 239-46 N1 - Jan JF - Infect Immun JO - Infection and immunity VL - 66 IS - 1 SN - 0019-9567 (Print) N1 - Temporal variation of the merozoite surface protein-2 gene of Plasmodium falciparum N1 - 9423864 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. United states N1 - eng KW - Alleles Amino Acid Sequence Animals *Antigens, Protozoan Chromosome Mapping DNA Primers/genetics DNA, Protozoan/analysis/genetics Epidemiology, Molecular Genome, Protozoan Humans Indonesia/epidemiology Malaria, Falciparum/blood/*epidemiology/*genetics Molecular Sequence Data Parasitemia/blood/epidemiology/genetics Plasmodium falciparum/*genetics Polymerase Chain Reaction Polymorphism, Genetic Protozoan Proteins/*genetics Repetitive Sequences, Nucleic Acid Sequence Alignment Sequence Analysis, DNA Sequence Homology, Amino Acid N2 - Extensive polymorphism of key parasite antigens is likely to hamper the effectiveness of subunit vaccines against Plasmodium falciparum infection. However, little is known about the extent of the antigenic repertoire of naturally circulating strains in different areas where malaria is endemic. To address this question, we conducted a study in which blood samples were collected from parasitemic individuals living within a small hamlet in Western Irian Jaya and subjected to PCR amplification using primers that would allow amplification of the gene encoding merozoite surface protein-2 (MSP2). We determined the nucleotide sequence of the amplified product and compared the deduced amino acid sequences to sequences obtained from samples collected in the same hamlet 29 months previously. MSP2 genes belonging to both major allelic families were observed at both time points. In the case of the FC27 MSP2 family, we observed that the majority of individuals were infected by parasites expressing the same form of MSP2. Infections with parasites expressing 3D7 MSP2 family alleles were more heterogeneous. No MSP2 alleles observed at the earlier time point were detectable at the later time point, either for the population as a whole or for individuals who were assayed at both time points. We examined a subset of the infected patients by using blood samples taken between the two major surveys. In no patients could we detect reinfection by a parasite expressing a previously encountered form of MSP2. Our results are consistent with the possibility that infection induces a form of strain-specific immune response against the MSP2 antigen that biases against reinfection by parasites bearing identical forms of MSP2. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9423864 ID - 152 ER - TY - JOUR AU - Coppel, R. L. AU - Cooke, B. M. AU - Magowan, C. AU - Narla, M. PY - 1998 TI - Malaria and the erythrocyte SP - 132-8 N1 - Mar JF - Curr Opin Hematol JO - Current opinion in hematology VL - 5 IS - 2 SN - 1065-6251 (Print) N1 - Malaria and the erythrocyte N1 - 9570706 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review United states N1 - eng KW - Animals Cell Adhesion Cytoskeleton/pathology Erythrocyte Membrane/pathology Erythrocytes/*parasitology/*pathology Humans Malaria/*blood/*parasitology/*pathology *Plasmodium falciparum N2 - In terms of global health, the most important disease involving human erythrocytes is infection by protozoan parasites of the genus Plasmodium, particularly Plasmodium falciparum. Our understanding of the complex processes of erythrocyte invasion, remodeling, and cytoadherence has advanced considerably over the past few years. Considerable advances have been made in identifying the players in each of these phenomena, although identification of the exact functional roles for many molecules is still missing. The cloning of the parasite adhesin, the development of a transfection system, and a series of new imaging and cell biology assays are recent achievements that promise to further our understanding not only of the pathogenesis of malaria, but also the functioning of erythrocytes. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9570706 ID - 150 ER - TY - JOUR AU - Coppel, R. L. AU - Brown, G. V. AU - Nussenzweig, V. PY - 1998 TI - Adhesive proteins of the malaria parasite SP - 472-81 N1 - Aug JF - Curr Opin Microbiol JO - Current opinion in microbiology VL - 1 IS - 4 SN - 1369-5274 (Print) N1 - Adhesive proteins of the malaria parasite N1 - 10066512 N1 - Ai32966/ai/niaid Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review England N1 - eng KW - Amino Acid Sequence Animals Cell Adhesion Cell Adhesion Molecules/genetics/*metabolism Erythrocytes/parasitology Liver/parasitology Malaria/*parasitology Molecular Sequence Data Plasmodium/genetics/*pathogenicity Protozoan Proteins/genetics/*metabolism N2 - Malaria infection of the host cells requires host-parasite recognition events mediated by adhesion and signaling molecules. Recent development of systems for stable transformation and targeted integration of exogenous DNA in malaria parasites provides a powerful tool to study the structure and function of Plasmodium attachment motifs, and their role in infection and disease. AD - Department of Microbiology, Monash University, Clayton, Victoria 3168, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10066512 ID - 141 ER - TY - JOUR AU - Cooke, B. M. AU - Nicoll, C. L. AU - Baruch, D. I. AU - Coppel, R. L. PY - 1998 TI - A recombinant peptide based on PfEMP-1 blocks and reverses adhesion of malaria-infected red blood cells to CD36 under flow SP - 83-90 N1 - Oct JF - Mol Microbiol JO - Molecular microbiology VL - 30 IS - 1 SN - 0950-382X (Print) N1 - A recombinant peptide based on PfEMP-1 blocks and reverses adhesion of malaria-infected red blood cells to CD36 under flow N1 - 9786187 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals Antigens, CD36/*metabolism Cell Adhesion Chondroitin Sulfates/metabolism Dose-Response Relationship, Drug Endothelium, Vascular/metabolism Erythrocytes/*parasitology/*physiology Hemorheology Humans Peptide Fragments/metabolism Plasmodium falciparum/*physiology Protozoan Proteins/*metabolism Recombinant Proteins/metabolism Species Specificity N2 - During falciparum malaria infection, severe complications ensue because parasitized red blood cells (PRBCs) adhere to endothelial cells and accumulate in the microvasculature. At the molecular level, adhesion is mediated by interaction of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP-1) on the PRBC surface with receptors on the surface of endothelial cells, including CD36. We have shown that a recombinant 179-residue subfragment of PfEMP-1 (rC1-2[1-179]), which encompasses the CD36-binding region, inhibits and reverses adhesion of PRBCs to CD36 under physiologically relevant flow conditions. rC1-2[1-179] inhibited adhesion in a concentration-dependent manner over the range 100 pM to 2 microM, with up to 99% of adhesion blocked at the highest concentration tested. The antiadhesive activity of rC1-2[1-179] was not strain specific and almost totally ablated adhesion of four different parasite lines. Furthermore, rC1-2[1-179] showed remarkable ability to progressively reverse adhesion when flowed over adherent PRBCs for 2h. The effect of rC1-2[1-179] was, however, specific for CD36-mediated adhesion and had no effect on adhesion mediated by CSA. Interference with binding of PRBCs to the vascular endothelium using rC1-2[1-179] or smaller organic mimetics may be a useful therapeutic approach to ameliorate severe complications of falciparum malaria. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9786187 ID - 146 ER - TY - JOUR AU - Van de Water, J. AU - Shimoda, S. AU - Niho, Y. AU - Coppel, R. AU - Ansari, A. AU - Gershwin, M. E. PY - 1997 TI - The role of T cells in primary biliary cirrhosis SP - 105-13 N1 - May JF - Semin Liver Dis JO - Seminars in liver disease VL - 17 IS - 2 SN - 0272-8087 (Print) N1 - The role of T cells in primary biliary cirrhosis N1 - 9170197 N1 - Journal Article Review United states N1 - eng KW - Antigen-Presenting Cells/immunology Autoantigens/analysis *Autoimmunity Epitopes/immunology Humans Liver Cirrhosis, Biliary/genetics/*immunology Major Histocompatibility Complex/genetics/immunology Mitochondria, Liver/immunology Phenotype Pyruvate Dehydrogenase Complex/immunology Receptors, Antigen, T-Cell/immunology T-Lymphocytes/*immunology N2 - While fervently studied by several laboratories, the role of T cells in the pathogenesis of primary biliary cirrhosis (PBC) still remains a mystery. The studies concerning cell phenotype, antigen specificity, and major histocompatibility complex (MHC)-T-cell receptor (TCR) interaction gathered thus far all address important aspects of this intriguing conundrum. However, the lack of an animal model and the genetic diversity of the human population with PBC make this task even more difficult. The possibilities regarding immune therapy resulting from such studies are of great importance. Future work concerning the T-cell epitopes--for both the pyruvate dehydrogenase complex (PDC), its related mitochondrial autoantigens, and any as yet unidentified PBC-specific autoantigens--may provide valuable information with regard to disease therapy. In addition, knowledge with regard to TCR usage and MHC association will help to clarify the pathogenic mechanisms of this enigmatic disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9170197 ID - 158 ER - TY - JOUR AU - Turchany, J. M. AU - Uibo, R. AU - Kivik, T. AU - Van de Water, J. AU - Prindiville, T. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1997 TI - A study of antimitochondrial antibodies in a random population in Estonia SP - 124-6 N1 - Jan JF - Am J Gastroenterol JO - The American journal of gastroenterology VL - 92 IS - 1 SN - 0002-9270 (Print) N1 - A study of antimitochondrial antibodies in a random population in Estonia N1 - 8995951 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Adolescent Adult Aged Aged, 80 and over Autoantibodies/*analysis Enzyme-Linked Immunosorbent Assay Estonia *Ethnic Groups Female Humans Immunoblotting Liver Cirrhosis, Biliary/immunology Male Middle Aged Mitochondria/*immunology Population Surveillance N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the spontaneous destruction of the small intrahepatic bile ducts. The hallmark serologic feature of PBC is the presence of high-titer antimitochondrial antibodies (AMA). Both the incidence and prevalence of PBC varies geographically; epidemiological data may provide valuable insight regarding the pathogenic mechanisms and etiology of disease. Thus far, the majority of studies on the occurrence of PBC and AMAs have been derived from autopsy, mortality figures, or hospital admission records. The numbers reported reflect only those patients with clinical disease. To address this issue, an adult population sample representing all age groups in the village of Karksi-Nuia in southern Estonia was selected for a study of AMA incidence. This village has unique features that make it ideal for such a study. First, the village is remote and a substantial number of families have lived in the area for generations. There is also a limited influx of new families into the village, therefore providing a limited genetic repertoire. In this unselected adult population, we examined AMA incidence by both immunoblot and ELISA, using native and recombinant antigens. Of the 1461 people studied, 13 (0.89%) were AMA positive. A similar frequency (0.96%) was found among 104 persons from a neighboring village, who subsequently joined the study. Our study suggests that the presence of AMA in Estonia is in agreement with the reported incidence of less than 1% AMA in a mixed hospital population. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California at Davis, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8995951 ID - 165 ER - TY - JOUR AU - Tsuneyama, K. AU - Van De Water, J. AU - Yamazaki, K. AU - Suzuki, K. AU - Sato, S. AU - Takeda, Y. AU - Ruebner, B. AU - Yost, B. A. AU - Nakanuma, Y. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1997 TI - Primary biliary cirrhosis an epithelitis: evidence of abnormal salivary gland immunohistochemistry SP - 23-31 JF - Autoimmunity JO - Autoimmunity VL - 26 IS - 1 SN - 0891-6934 (Print) N1 - Primary biliary cirrhosis an epithelitis: evidence of abnormal salivary gland immunohistochemistry N1 - 9556352 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Switzerland N1 - eng KW - Animals Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/immunology Fluorescent Antibody Technique, Indirect Humans Liver Cirrhosis, Biliary/*immunology/pathology Mice Pyruvate Dehydrogenase Complex/analysis Salivary Glands/*immunology/pathology Sjogren's Syndrome/*immunology/pathology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease of unknown etiology. Nearly 93% of patients with PBC exhibit evidence of focal sialoadenitis. In an earlier study, we reported evidence of aberrant expression of PDC-E2, or a mimeotope, in the salivary glands of patients with PBC that had Sjogren's syndrome. At the time of the previous study, data was not yet available regarding patients with PBC without sicca complaints. Therefore, to investigate the extent of salivary gland involvement in PBC, we collected lip biopsy sections from 9 PBC patients diagnosed as PBC by liver biopsy, without clinical or histologic features of Sjogren's syndrome and 9 PBC patients with established Sjogren's syndrome. Using immunohistochemical staining with both a murine monoclonal antibody. C355.1, and a human combinatorial antibody, SP4, we examined the ducts of these salivary glands for the presence of the characteristic aberrant staining pattern found in patients with PBC. We report that 6/9 PBC patients fulfilling established Sjogren's syndrome criteria and 6/9 PBC patients lacking features of Sjogren's syndrome showed intense staining of the ductal epithelial cells of the salivary gland. These data suggest that the PBC-specific antigen recognized by C355.1 and SP4 in bile duct epithelial cells is expressed aberrantly in the salivary gland in 66% of patients with PBC, independent of Sjogren's syndrome. This finding suggests a common disease process in these two tissues. Further, expression of this molecule may be an early marker of salivary gland involvement in patients with PBC. AD - Department of Pathology, Kanazawa University School of Medicine, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9556352 ID - 164 ER - TY - JOUR AU - Nishio, A. AU - Van de Water, J. AU - Leung, P. S. AU - Joplin, R. AU - Neuberger, J. M. AU - Lake, J. AU - Bjorkland, A. AU - Totterman, T. H. AU - Peters, M. AU - Worman, H. J. AU - Ansari, A. A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1997 TI - Comparative studies of antimitochondrial autoantibodies in sera and bile in primary biliary cirrhosis SP - 1085-9 N1 - May JF - Hepatology JO - Hepatology (Baltimore, Md VL - 25 IS - 5 SN - 0270-9139 (Print) N1 - Comparative studies of antimitochondrial autoantibodies in sera and bile in primary biliary cirrhosis N1 - 9141421 N1 - Dk39558/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Autoantibodies/*blood Autoantigens/analysis/immunology Autoimmunity Bile/*immunology Humans Liver Cirrhosis, Biliary/blood/*immunology Mitochondria/*immunology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by destruction of intrahepatic bile ducts. Although the pathogenesis of this disease is still unknown, high titers of antimitochondrial autoantibodies (AMA) have long been recognized in patient sera. However, little is known about the presence of AMA in bile. In this study, we investigated bile and sera from patients with PBC and healthy controls for the presence of AMA and mitochondrial autoantigens. AMA were detected in the bile of 17 of 19 patients (89.4%) with PBC; they were specifically directed against the pyruvate dehydrogenase complex (PDC-E2) in 15 of 19 patients (78.9%), to the branched-chain 2-oxo-acid dehydrogenase complex E2 (BCOADC-E2) in 6 of 19 patients (31.6%), and to the 2-oxoglutarate dehydrogenase complex E2 (OGDC-E2) in 1 of 19 patients (5.3%). In a comparative study of sera from the same patients, anti-PDC-E2 antibodies were found in 19 of 19 patients (100%), anti-BCOADC in 9 of 19 patients (47.3%), and anti-OGDC-E2 in 4 of 19 patients (21.1%) patients. AMA in bile were always found together with antibodies of corresponding specificities in the serum from the same patient. Immunoglobulin (Ig)A AMA were found in the bile of 9 of 19 patients (47.7%) with PBC; they were specifically directed against PDC-E2 in 8 of 19 patients (42.1%) and to BCOADC in 2 of 19 patients (10.5%). Epitope mapping of IgA anti-PDC-E2 antibodies indicated that, like serum autoantibodies, the immunodominant epitope is directed against the inner lipoyl domain of PDC-E2. The prevalence and antigen reactivity of IgA AMA in sera correlated completely with IgA AMA in bile. Autoantibodies against nuclear envelope pore proteins (gp210) were found in 1 of 8 (12.5%) sera of patients with PBC, but not in bile. Furthermore, and of particular interest, we detected the autoantigens, PDC-E2, OGDC-E2, and BCOADC-E2, in the bile of 12 of 19 patients (63.2%), 9 of 19 patients (47.4%), and 9 of 19 patients (47.4%), respectively; PDC-E2 was found in only 1 of 17 (5.9%) disease controls. Although the presence of AMA in bile may merely reflect the presence of these antibodies in sera, the simultaneous detection of mitochondrial autoantigens in bile suggests an increase of mitochondrial autoantigens at inflammatory sites. Such autoantigens, coupled with AMA, may augment the local immune response and disease progression. AD - Division of Rheumatology, Allergy, and Clinical Immunology, University of California at Davis, School of Medicine, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9141421 ID - 159 ER - TY - JOUR AU - Marshall, V. M. AU - Silva, A. AU - Foley, M. AU - Cranmer, S. AU - Wang, L. AU - McColl, D. J. AU - Kemp, D. J. AU - Coppel, R. L. PY - 1997 TI - A second merozoite surface protein (MSP-4) of Plasmodium falciparum that contains an epidermal growth factor-like domain SP - 4460-7 N1 - Nov JF - Infect Immun JO - Infection and immunity VL - 65 IS - 11 SN - 0019-9567 (Print) N1 - A second merozoite surface protein (MSP-4) of Plasmodium falciparum that contains an epidermal growth factor-like domain N1 - 9353020 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Surface/chemistry Base Sequence DNA, Complementary/isolation & purification Epidermal Growth Factor/*chemistry Glycosylphosphatidylinositols/analysis Molecular Sequence Data Plasmodium falciparum/*chemistry Protozoan Proteins/*chemistry/genetics N2 - Merozoite surface proteins of Plasmodium falciparum play a critical role in the invasion of human erythrocytes by the malaria parasite. Here we describe the identification of a novel protein with a molecular mass of 40 kDa that is found on the merozoite surface of P. falciparum. We call this protein merozoite surface protein 4 (MSP-4). Evidence for the surface location of MSP-4 includes (i) a staining pattern that is consistent with merozoite surface location in indirect immunofluorescent studies of cultured parasites, (ii) localization of MSP-4 in the detergent phase in Triton X-114 partitioning studies, and (iii) nucleotide sequencing studies which predict the presence of an N-terminal signal sequence and a hydrophobic C-terminal sequence in the protein. Immunoprecipitation studies of biosynthetically labelled parasites with [3H] myristic acid indicated that MSP-4 is anchored on the merozoite surface by a glycosylphosphatidylinositol moiety. Of considerable interest is the presence of a single epidermal growth factor-like domain at the C terminus of the MSP-4 protein, making it the second protein with such a structure to be found on the merozoite surface. AD - The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. vmarshal@wehi.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9353020 ID - 153 ER - TY - JOUR AU - Marshall, V. M. AU - Coppel, R. L. PY - 1997 TI - Characterisation of the gene encoding adenylosuccinate lyase of Plasmodium falciparum SP - 237-41 N1 - Sep JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 88 IS - 1-2 SN - 0166-6851 (Print) N1 - Characterisation of the gene encoding adenylosuccinate lyase of Plasmodium falciparum N1 - 9274883 N1 - Journal Article Netherlands N1 - eng KW - Adenylosuccinate Lyase/*genetics Amino Acid Sequence Animals *Antigens, Protozoan *Genes, Protozoan Humans Molecular Sequence Data Plasmodium falciparum/*enzymology/*genetics Protozoan Proteins/genetics Sequence Homology, Amino Acid Species Specificity AD - The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia. vmarshal@wehi.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9274883 ID - 154 ER - TY - JOUR AU - Magowan, C. AU - Brown, J. T. AU - Liang, J. AU - Heck, J. AU - Coppel, R. L. AU - Mohandas, N. AU - Meyer-Ilse, W. PY - 1997 TI - Intracellular structures of normal and aberrant Plasmodium falciparum malaria parasites imaged by soft x-ray microscopy SP - 6222-7 N1 - Jun 10 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 94 IS - 12 SN - 0027-8424 (Print) N1 - Intracellular structures of normal and aberrant Plasmodium falciparum malaria parasites imaged by soft x-ray microscopy N1 - 9177198 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Cysteine Proteinase Inhibitors/pharmacology *Cytoskeletal Proteins Electron Probe Microanalysis/methods Erythrocyte Membrane/physiology Erythrocytes/*parasitology Erythrocytes, Abnormal/*parasitology Humans Membrane Proteins/deficiency/physiology *Neuropeptides Plasmodium falciparum/drug effects/*ultrastructure N2 - Soft x-ray microscopy is a novel approach for investigation of intracellular organisms and subcellular structures with high spatial resolution. We used x-ray microscopy to investigate structural development of Plasmodium falciparum malaria parasites in normal and genetically abnormal erythrocytes and in infected erythrocytes treated with cysteine protease inhibitors. Investigations in normal red blood cells enabled us to recognize anomalies in parasite structures resulting from growth under unfavorable conditions. X-ray microscopy facilitated detection of newly elaborated structures in the cytosol of fixed, unstained, intact erythrocytes, redistribution of mass (carbon) in infected erythrocytes, and aberrant parasite morphology. In cysteine protease inhibitor-treated, infected erythrocytes, high concentrations of material were detected in abnormal digestive vacuoles and aggregated at the parasite plasma membrane. We have demonstrated that an abnormal host erythrocyte skeleton affects structural development of parasites and that this aberrant development can be detected in the following generation when parasites from protein 4.1-deficient red blood cells infect normal erythrocytes. This work extends our current understanding of the relationship between the host erythrocyte membrane and the intraerythrocytic malaria parasite by demonstrating for the first time that constituents of the erythrocyte membrane play a role in normal parasite structural development. AD - Life Sciences Division, Lawrence Berkeley National Laboratory, One Cyclotron Road, Berkeley, CA 94720, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9177198 ID - 156 ER - TY - JOUR AU - Leung, P. S. AU - Coppel, R. L. AU - Ansari, A. AU - Munoz, S. AU - Gershwin, M. E. PY - 1997 TI - Antimitochondrial antibodies in primary biliary cirrhosis SP - 61-9 N1 - Feb JF - Semin Liver Dis JO - Seminars in liver disease VL - 17 IS - 1 SN - 0272-8087 (Print) N1 - Antimitochondrial antibodies in primary biliary cirrhosis N1 - 9089911 N1 - Ai 31585/ai/niaid Dk 99588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Autoantibodies/*analysis/genetics Autoantigens/analysis/genetics Autoimmune Diseases/*immunology Bile Ducts, Intrahepatic/immunology Biochemistry Cloning, Molecular Epithelium/immunology Epitopes Genes, Immunoglobulin/genetics Humans Immunohistochemistry Isoenzymes/analysis/genetics/immunology Ketone Oxidoreductases/analysis/genetics/immunology Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/*immunology Multienzyme Complexes/analysis/genetics/immunology Thioctic Acid/analysis/genetics/immunology N2 - In the last decade, the cloning and biochemical identification of mitochondrial autoantigens in primary biliary cirrhosis (PBC) as members of the 2-oxoacid dehydrogenase complex has greatly advanced the detection of antimitochondrial antibodies (AMA) and the understanding of the immunobiology of the disease. Here, we discuss the methods of detecting AMA and its isotypes, methods of epitope mapping, and using these methods in PBC liver immunohistochemistry and Ig gene usage. Increasing evidence, including the specific association of AMA with PBC, the unique similar but noncross-reactive conformational epitope of the lipoyl domains of the mitochondrial autoantigens, the specific binding of anti-PDC-E2 monoclonal antibodies and human combinatorial antibodies derived from PBC patients to the apical area of bile duct epithelial cells in PBC livers, and Ig gene usage of AMA, suggests that AMA is not an epiphenomenon of the disease but plays a significant role in the pathogenesis of PBC. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9089911 ID - 163 ER - TY - JOUR AU - Kun, J. F. AU - Hibbs, A. R. AU - Saul, A. AU - McColl, D. J. AU - Coppel, R. L. AU - Anders, R. F. PY - 1997 TI - A putative Plasmodium falciparum exported serine/threonine protein kinase SP - 41-51 N1 - Mar JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 85 IS - 1 SN - 0166-6851 (Print) N1 - A putative Plasmodium falciparum exported serine/threonine protein kinase N1 - 9108547 N1 - Dk 32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Surface/isolation & purification Cell Compartmentation Cloning, Molecular Erythrocyte Membrane/ultrastructure Escherichia coli/genetics Fluorescent Antibody Technique *Genes, Protozoan Microscopy, Immunoelectron Molecular Sequence Data Plasmodium falciparum/enzymology/*genetics/ultrastructure Protein-Serine-Threonine Kinases/*genetics/secretion Protozoan Proteins/*genetics/isolation & purification/secretion Recombinant Fusion Proteins/biosynthesis Sequence Analysis, DNA N2 - An 8kb gene coding for a putative serine/threonine protein kinase from Plasmodium falciparum has been cloned and sequenced. It is arranged in two exons: exon I is 2 kb and exon II is 5.6 kb. The gene codes for a large protein of 2510 amino acids. Antibodies raised against a fusion protein were used to localize the putative kinase. By immunofluorescence microscopy, it was found in the cytoplasm of infected red cells. By immunoelectron microscopy it was associated with membranous structures in the red cell and with the red cell membrane, particularly at parasite-induced knobs. This is the first putative protein kinase of P. falciparum to be exported from the parasite into its host cell. AD - Australian Centre for International and Tropical Health and Nutrition, Royal Brisbane Hospital, Australia Qld. juergen.kun@uni-tuebingen.de UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9108547 ID - 162 ER - TY - JOUR AU - Harada, K. AU - Van de Water, J. AU - Leung, P. S. AU - Coppel, R. L. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 1997 TI - In situ nucleic acid hybridization of pyruvate dehydrogenase complex-E2 in primary biliary cirrhosis: pyruvate dehydrogenase complex-E2 messenger RNA is expressed in hepatocytes but not in biliary epithelium SP - 27-32 N1 - Jan JF - Hepatology JO - Hepatology (Baltimore, Md VL - 25 IS - 1 SN - 0270-9139 (Print) N1 - In situ nucleic acid hybridization of pyruvate dehydrogenase complex-E2 in primary biliary cirrhosis: pyruvate dehydrogenase complex-E2 messenger RNA is expressed in hepatocytes but not in biliary epithelium N1 - 8985260 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Bile Ducts/*metabolism Humans Liver/*metabolism Liver Cirrhosis, Biliary/*enzymology *Nucleic Acid Hybridization Pyruvate Dehydrogenase Complex/*genetics RNA, Messenger/*analysis N2 - Pyruvate dehydrogenase-E2, or a cross-reactive molecule, has been shown by a variety of immunohistochemical methods to be present in increased amounts in biliary epithelial cells (BEC) in primary biliary cirrhosis (PBC). In this study, to further understand the nature of the immunoreactive molecule in BEC, we examined the expression of pyruvate dehydrogenase complex-E2 (PDC-E2) messenger RNA (mRNA) and PDC-E2 protein in sections of livers from patients and controls to help identify the molecule found in BEC. We performed in situ hybridization using an antisense probe against the major epitope of PDC-E2. The data were very striking and suggested that there was no increased production of PDC-E2 in BEC. For example, in livers from patients with PBC, PDC-E2 mRNA was found in periportal hepatocytes in 16 of 17 cases (94%). In contrast, interlobular bile ducts and septal bile ducts had detectable levels of PDC-E2 mRNA in only 1 of 17 (6%) and 3 of 8 (38%) cases, respectively. Interestingly, proliferating bile ductules contained detectable levels of mRNA in 12 of 15 cases (80%). In control liver, periportal hepatocytes were positive in 15 of 17 cases (88%). Interlobular bile ducts, septal bile ducts, and proliferating bile ductules expressed mRNA signals in 4 of 17 (24%), 2 of 10 (20%), and 14 of 16 (88%), respectively. When formalin-fixed, paraffin-embedded sections were examined by immunohistochemical staining with anti-PDC-E2 monoclonal antibody (mAb) C355.1, the interlobular bile ducts showed typical aberrant apical staining in all 10 PBC cases, but 0 of 9 liver controls. Periportal hepatocytes, proliferating bile ductules and infiltrating mononuclear cells stained with C355.1 but in a characteristic mitochondrial staining pattern. The presence of a PDC-E2-like molecule recognized by C355.1 is not reflected by the expression levels of PDC-E2 mRNA in the BEC of patients with PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8985260 ID - 166 ER - TY - JOUR AU - Harada, K. AU - Van de Water, J. AU - Leung, P. S. AU - Coppel, R. L. AU - Ansari, A. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 1997 TI - In situ nucleic acid hybridization of cytokines in primary biliary cirrhosis: predominance of the Th1 subset SP - 791-6 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 25 IS - 4 SN - 0270-9139 (Print) N1 - In situ nucleic acid hybridization of cytokines in primary biliary cirrhosis: predominance of the Th1 subset N1 - 9096578 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Actins/genetics Base Sequence Case-Control Studies DNA Primers/genetics Fructose-Bisphosphate Aldolase/genetics Gene Expression Humans In Situ Hybridization Interferon Type II/*genetics Interleukin-4/*genetics Liver/immunology/pathology Liver Cirrhosis, Biliary/*genetics/*immunology/pathology RNA, Messenger/genetics/metabolism Th1 Cells/*immunology/pathology Th2 Cells/immunology/pathology N2 - Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by destruction of the intrahepatic bile ducts. It is generally believed that cellular immune mechanisms, particularly involving T cells, result in this bile duct damage. The relative strength of Th1 and Th2 responses has recently been proposed to be an important factor in the pathophysiology of various autoimmune diseases. In this study, we have attempted to identify the Th subset balance in PBC, by detection of cytokines specific to the two T-cell subsets, i.e., interferon gamma (IFN-gamma) for Th1 cells and interleukin-4 (IL-4) for Th2 cells. We analyzed IFN-gamma and IL-4 messenger RNA (mRNA) positive cells in liver sections from 18 patients with PBC and 35 disease controls including chronic active hepatitis C, extrahepatic biliary obstruction (EBO), and normal liver, using nonisotopic in situ hybridization and immunohistochemistry. Mononuclear cells expressing IFN-gamma and IL-4 mRNA were aggregated in inflamed portal tracts in PBC livers, but were rarely present in extrahepatic biliary obstruction, alcoholic fibrosis, or normal liver sections. The IFN-gamma and IL-4 mRNA positive cells in PBC livers were detected in significantly higher numbers than in control livers (P < .01). Moreover, IFN-gamma mRNA expression was more commonly detected than IL-4 expression in PBC livers, and the levels of IFN-gamma mRNA expression were highly correlated with the degree of portal inflammatory activity. IFN-gamma mRNA-positive cells were detected primarily around damaged bile ducts that were surrounded by lymphoid aggregates. The data indicate that Th1 cells are the more prominent T-cell subset in the lymphoid infiltrates in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9096578 ID - 161 ER - TY - JOUR AU - Cranmer, S. L. AU - Magowan, C. AU - Liang, J. AU - Coppel, R. L. AU - Cooke, B. M. PY - 1997 TI - An alternative to serum for cultivation of Plasmodium falciparum in vitro SP - 363-5 N1 - May-Jun JF - Trans R Soc Trop Med Hyg JO - Transactions of the Royal Society of Tropical Medicine and Hygiene VL - 91 IS - 3 SN - 0035-9203 (Print) N1 - An alternative to serum for cultivation of Plasmodium falciparum in vitro N1 - 9231219 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Animals *Culture Media Parasitology/methods Plasmodium falciparum/*growth & development *Serum Albumin, Bovine Time Factors AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9231219 ID - 157 ER - TY - JOUR AU - Crabb, B. S. AU - Cooke, B. M. AU - Reeder, J. C. AU - Waller, R. F. AU - Caruana, S. R. AU - Davern, K. M. AU - Wickham, M. E. AU - Brown, G. V. AU - Coppel, R. L. AU - Cowman, A. F. PY - 1997 TI - Targeted gene disruption shows that knobs enable malaria-infected red cells to cytoadhere under physiological shear stress SP - 287-96 N1 - Apr 18 JF - Cell JO - Cell VL - 89 IS - 2 SN - 0092-8674 (Print) N1 - Targeted gene disruption shows that knobs enable malaria-infected red cells to cytoadhere under physiological shear stress N1 - 9108483 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Antigens, CD36/metabolism Blood Platelets/metabolism Blood Proteins/analysis Cell Adhesion/*physiology Erythrocyte Membrane/chemistry/ultrastructure Erythrocytes/*cytology/*parasitology Gene Expression Molecular Sequence Data Mutagenesis Peptides/genetics/*physiology Plasmodium falciparum/*physiology Protozoan Proteins/analysis Stress, Mechanical Transfection N2 - Knobs at the surface of erythrocytes infected with Plasmodium falciparum have been proposed to be important in adherence of these cells to the vascular endothelium. This structure contains the knob-associated histidine-rich protein (KAHRP) and the adhesion receptor P. falciparum erythrocyte membrane protein 1. We have disrupted the gene encoding KAHRP and show that it is essential for knob formation. Knob-transfectants adhere to CD36 in static assays; when tested under flow conditions that mimic those of postcapillary venules, however, the binding to CD36 was dramatically reduced. These data suggest that knobs on P. falciparum-infected erythrocytes exert an important influence on adherence of parasitized-erythrocytes to microvascular endothelium, an important process in the pathogenesis of P. falciparum infections. AD - The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9108483 ID - 160 ER - TY - JOUR AU - Bennett, B. J. AU - Mohandas, N. AU - Coppel, R. L. PY - 1997 TI - Defining the minimal domain of the Plasmodium falciparum protein MESA involved in the interaction with the red cell membrane skeletal protein 4.1 SP - 15299-306 N1 - Jun 13 JF - J Biol Chem JO - The Journal of biological chemistry VL - 272 IS - 24 SN - 0021-9258 (Print) N1 - Defining the minimal domain of the Plasmodium falciparum protein MESA involved in the interaction with the red cell membrane skeletal protein 4.1 N1 - 9182557 N1 - Dk32094-10/dk/niddk In Vitro Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals *Cytoskeletal Proteins Cytoskeleton/metabolism Erythrocytes/*metabolism Humans Membrane Proteins/*metabolism *Neuropeptides Plasmodium falciparum/*metabolism Protozoan Proteins/*metabolism N2 - During part of its life cycle, the protozoan parasite Plasmodium falciparum lives within the human red blood cell and modifies both the structural and functional properties of the red cell. It does this by synthesizing a number of polypeptides that it transports into the red cell cytoplasm and to the red cell membrane. One of these transported proteins, MESA (mature parasite-infected erythrocyte surface antigen), is anchored to the red cell membrane by noncovalent interaction with erythrocyte protein 4.1. We have utilized a combination of in vitro transcription and translation and a membrane binding assay to identify the protein sequence involved in anchoring MESA to the membrane. Labeled fragments of different regions of the MESA protein were evaluated for their ability to bind to inside-out vesicle membrane preparations of human red cells. Binding was dependent on the presence of red cell membrane proteins and was abolished either by trypsin treatment or by selective depletion of membrane proteins. Binding was specific and could be inhibited by the addition of competing protein, with an IC50 of (6.3 +/- 1.2) x 10(-7) M, indicative of a moderate affinity interaction. Fractionation studies demonstrated that binding fragments interacted most efficiently with membrane protein fractions that had been enriched in protein 4.1. Binding inhibition experiments using synthetic peptides identified the binding domain of MESA for protein 4.1 as a 19-residue sequence near the amino terminus of MESA, a region capable of forming an amphipathic helix. AD - Walter and Eliza Hall Institute of Medical Research, Victoria 3050, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9182557 ID - 155 ER - TY - JOUR AU - Yip, T. T. AU - Van de Water, J. AU - Gershwin, M. E. AU - Coppel, R. L. AU - Hutchens, T. W. PY - 1996 TI - Cryptic antigenic determinants on the extracellular pyruvate dehydrogenase complex/mimeotope found in primary biliary cirrhosis. A probe by affinity mass spectrometry SP - 32825-33 N1 - Dec 20 JF - J Biol Chem JO - The Journal of biological chemistry VL - 271 IS - 51 SN - 0021-9258 (Print) N1 - Cryptic antigenic determinants on the extracellular pyruvate dehydrogenase complex/mimeotope found in primary biliary cirrhosis. A probe by affinity mass spectrometry N1 - 8955120 N1 - 39588/phs 50977/phs R41gm 51658/gm/nigms Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Antibodies, Monoclonal/*diagnostic use/immunology Autoantigens/*immunology Epitopes Humans Liver/immunology Liver Cirrhosis, Biliary/*immunology Mass Spectrometry/*methods Mitochondria/immunology Pyruvate Dehydrogenase Complex/*immunology N2 - Affinity mass spectrometry (AMS) was used to evaluate the structural diversity of the E2 component of pyruvate dehydrogenase complex (PDC) in normal and diseased liver cells, including those from patients with the autoimmune disease primary biliary cirrhosis (PBC). Two different antibodies to PDC-E2, the immunodominant mitochondrial autoantigen in patients with PBC, were used. AMS was performed directly on frozen liver sections and purified bile duct epithelial cells. Mass spectrometric signals associated with the molecular recognition of PBC-specific antigenic determinants were enhanced by an in situ enzyme-linked signal amplification process. Samples from patients with PBC gave strong positive signals for the antigen(s) recognized by the monoclonal antibody C355.1. Conversely, tissues from normal and disease controls showed only a minimal signal. AMS was used to identify specific antigenic determinants within the E2 component of PDC for comparison with unknown antigenic determinants observed by affinity capture with C355.1 monoclonal antibody from PBC samples. PDC components bound to C355.1 were mapped and identified by mass before dissociation from the E2 component. A similar approach was used to identify unknown antigenic determinants associated with PBC. We believe AMS may be an important new approach with wide application to the identification of molecules associated with a number of disease states. AD - Department of Food Science and Technology, Allergy and Clinical Immunology, University of California Davis, School of Medicine, Davis, California 95616, USA. megershwin@ucdavis.edu UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8955120 ID - 167 ER - TY - JOUR AU - Van de Water, J. AU - Gerson, L. B. AU - Ferrell, L. D. AU - Lake, J. R. AU - Coppel, R. L. AU - Batts, K. P. AU - Wiesner, R. H. AU - Gershwin, M. E. PY - 1996 TI - Immunohistochemical evidence of disease recurrence after liver transplantation for primary biliary cirrhosis SP - 1079-84 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 24 IS - 5 SN - 0270-9139 (Print) N1 - Immunohistochemical evidence of disease recurrence after liver transplantation for primary biliary cirrhosis N1 - 8903379 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Alkaline Phosphatase/blood Animals Antibodies, Monoclonal/immunology Female Humans Immunohistochemistry Liver Cirrhosis, Biliary/enzymology/*surgery *Liver Transplantation Mice Middle Aged Pyruvate Dehydrogenase Complex/*analysis Recurrence N2 - Whether primary biliary cirrhosis (PBC) recurs after liver transplantation has remained an interesting and controversial issue; rejection, viral hepatitis, and drug effects all may mimic recurrent PBC histologically and biochemically. Furthermore, reliable clinical criteria for PBC recurrence are lacking. In this study, the issue of disease recurrence using a well-characterized monoclonal antibody (MAb), C355.1, that reacts with the immunodominant mitochondrial autoantigen of PBC (pyruvate dehydrogenase complex [PDC-E2]) was addressed. When used in an immunohistochemical assay, C355.1 produces intense apical staining of bile duct epithelium specifically in liver sections of patients with PBC and may be the earliest known marker of PBC. Immunohistochemical and histological analysis of serial liver biopsy specimens of 67 patients pre- and post-orthotopic liver transplantation (OLT), including 38 patients with PBC and 29 non-PBC liver disease controls, was performed. Sections were stained with MAb C355.1 or the control MAb C315 and analyzed to determine whether there was a recurrence of apical reactivity in the bile ducts of the posttransplantation biopsy specimens. The immunohistochemical staining was correlated with the histological findings and serum biochemistries at the time of the biopsy. Our data indicate that a significant number of patients who underwent transplantation for PBC (28 of 38) but not controls (0 of 29) develop a staining pattern of liver bile duct epithelium with MAb C355.1 that is indistinguishable from the pretransplantation pattern. Of the 28 patients with this apical staining pattern, 8 were characterized histologically as possible recurrent PBC, 2 as chronic rejection, 2 as acute rejection, 9 as nonspecific changes, 4 as normal or near normal, and 3 had other histological changes. Only 50% of the patients with apical C355.1 staining had liver enzyme levels suggestive of cholestasis. Thus, there appears to be immunohistochemical evidence that supports the concept of recurrence of PBC after OLT. The appearance of biliary epithelial abnormalities before the clinical appearance of disease is important not only for liver transplantation but also for understanding the natural history of PBC. AD - Department of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California Davis, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8903379 ID - 171 ER - TY - JOUR AU - Moteki, S. AU - Leung, P. S. AU - Dickson, E. R. AU - Van Thiel, D. H. AU - Galperin, C. AU - Buch, T. AU - Alarcon-Segovia, D. AU - Kershenobich, D. AU - Kawano, K. AU - Coppel, R. L. AU - et al. PY - 1996 TI - Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex SP - 436-44 N1 - Mar JF - Hepatology JO - Hepatology (Baltimore, Md VL - 23 IS - 3 SN - 0270-9139 (Print) N1 - Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex N1 - 8617422 N1 - Ai 31585/ai/niaid Dk 39558/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Antigen-Antibody Reactions Autoantibodies/blood/*immunology Autoantigens/*immunology Base Sequence Epitopes/*immunology Humans Immunoblotting Ketoglutarate Dehydrogenase Complex/genetics/*immunology Liver Cirrhosis, Biliary/enzymology/*immunology Mitochondria/immunology Molecular Sequence Data Rats Recombinant Fusion Proteins/immunology N2 - Five different target mitochondrial autoantigens recognized by sera from patients with primary biliary cirrhosis (PBC) have been identified as subunits of the following 2-oxo acid dehydrogenase complexes: the pyruvate dehydrogenase complex (PDC), the branched chain 2-oxo acid dehydrogenase complex (BCOADC), and the 2-oxoglutarate dehydrogenase complex (OGDC). Unlike the E2 subunits of PDC (PDC-E2) and BCOADC (BCOADC-E2), the E2 subunits of OGDC (OGDC-E2) reactivity of PBC sera and the reactive epitope of OGDC-D2 have not hitherto been studied in detail. In this report, we took advantage of a recombinant fusion protein for OGDC-E2 to address these issues. Eighty of 268 (29.9%) PBC patient sera but none of the 45 controls reacted with recombinant OGDC-E2. The recombinant OGDC-E2 was judged to express the immunodominant epitope, because when sera from patients with PBC were preabsorbed with the recombinant fusion protein, such sera were depleted of reactivity against 48 kD OGDC-E2 when probed on beef heart mitochondria (BHM) but retained reactivity toward PDC-E2 and/or BCOADC-E2. Furthermore, affinity-purified PBC sera against recombinant OGDC-E2 reacted only with native OGDC-E2 and not with any other enzyme components of the 2-oxo acid dehydrogenase complex. Antimitochondrial autoantibodies (AMA) against OGDC-E2 included immunoglobulin (Ig)G2, IgG3 and IgM and the relative titers were as follows: IgG2 > IgG3 > IgM. Finally, using overlapping recombinant polypeptides, it was determined that a minimum of 81 amino acids (residues 67-147) corresponding to the lipoyl domain of OGDC-E2 are necessary for reactivity, suggesting that a conformational autoepitope is recognized by AMA. These data suggest that each of the 2-oxo acid dehydrogenase enzymes has distinct antigenicity despite their similarities in structure and function. The availability of recombinant OGDC-E2 autoantigen will allow the design of additional studies to further our understanding of the role of mitochondrial autoantigens in the pathogenesis of PBC. AD - Division of Rheumatology, Allergy, and Clinical Immunology, School of Medicine, University of California Davis, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8617422 ID - 178 ER - TY - JOUR AU - Moteki, S. AU - Leung, P. S. AU - Coppel, R. L. AU - Dickson, E. R. AU - Kaplan, M. M. AU - Munoz, S. AU - Gershwin, M. E. PY - 1996 TI - Use of a designer triple expression hybrid clone for three different lipoyl domain for the detection of antimitochondrial autoantibodies SP - 97-103 N1 - Jul JF - Hepatology JO - Hepatology (Baltimore, Md VL - 24 IS - 1 SN - 0270-9139 (Print) N1 - Use of a designer triple expression hybrid clone for three different lipoyl domain for the detection of antimitochondrial autoantibodies N1 - 8707289 N1 - Comparative Study Journal Article United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Animals Autoantibodies/*blood Autoantigens/*immunology Base Sequence Binding Sites Cattle Cloning, Molecular/*methods DNA Primers Diagnosis, Differential Enzyme-Linked Immunosorbent Assay/methods Epitopes/*analysis Fluorescent Antibody Technique Humans Ketoglutarate Dehydrogenase Complex/*immunology Ketone Oxidoreductases/*immunology Liver Cirrhosis, Biliary/blood/*diagnosis/immunology Mitochondria/*immunology Molecular Sequence Data Multienzyme Complexes/*immunology Polymerase Chain Reaction/*methods Protein Hybridization Pyruvate Dehydrogenase Complex/*immunology Rats Recombinant Fusion Proteins/immunology Reference Values Reproducibility of Results Restriction Mapping Sensitivity and Specificity Thioctic Acid/metabolism N2 - The detection of antimitochondrial autoantibodies (AMAs) is critical in the diagnosis of primary biliary cirrhosis (PBC). However, conventional laboratory assays to detect AMA are dependent on the time-consuming method of immunofluorescence microscopy, a method often plagued by problems of nonspecificity. AMAs react against mitochondrial autoantigens including the E2 components of the pyruvate dehydrogenase complex (PDC-E2), the branched-chain 2-oxo-acid dehydrogenase complex (BCOADC-E2), and the 2-oxo-glutarate dehydrogenase complex (OGDC-E2). Interestingly, the immunodominant epitopes of PDC-E2, BCOADC-E2, and OGDC-E2 are all conformational lipoate binding sites, but antibodies against them do not cross-react. Although 80% to 90% of sera from patients with PBC react to PDC-E2, approximately 10% patients with PBC react only to BCOADC-E2 and/or OGDC-E2. We have taken advantage of our epitope-mapping studies of the E2 components of PDC, BCOADC, and OGDC, and constructed a "designer" hybrid clone, designated as pML-MIT3, that coexpresses the immunodominant epitopes within the three distinct lipoyl domains. We examined a total of 321 sera, including 186 sera from patients with PBC, to test the immunoreactivity of pMIT3. Of 186 sera from patients with PBC, 152 sera (81.7%) reacted with recombinant fusion protein of PDC-E2, whereas 171 sera (91.9%) showed positive reactivities when probed by immunoblotting against the recombinant fusion protein expressed from the pML-MIT3 clone. Of 34 PBC sera that did not react with recombinant PDC-E2, 18 sera contained BCOADC-E2-specific AMA and 1 serum possessed only OGDC-E2-specific AMA. We also developed an enzyme-linked immunosorbent assay (ELISA), using affinity-purified recombinant fusion protein of pML-MIT3 clone as the antigen source, to quantify specific AMAs in patients with PBC. None of the 135 control sera from patients with primary sclerosing cholangitis (PSC), chronic autoimmune hepatitis (CAH), systemic lupus erythematosus (SLE), or healthy volunteers showed significant reactivity against pML-MIT3 recombinant fusion protein in the ELISA assay. Our results indicate that an ELISA using recombinant, cloned autoantigen of pML-MIT3 is a powerful and very specific method for the detection of AMA. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California Davis, CA 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8707289 ID - 175 ER - TY - JOUR AU - Marshall, V. M. AU - Zhang, L. AU - Anders, R. F. AU - Coppel, R. L. PY - 1996 TI - Diversity of the vaccine candidate AMA-1 of Plasmodium falciparum SP - 109-13 N1 - Apr JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 77 IS - 1 SN - 0166-6851 (Print) N1 - Diversity of the vaccine candidate AMA-1 of Plasmodium falciparum N1 - 8784778 N1 - Comparative Study Journal Article Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/biosynthesis/chemistry/immunology Base Sequence DNA Primers Membrane Proteins/biosynthesis/chemistry/*immunology Molecular Sequence Data Plasmodium falciparum/genetics/*immunology/isolation & purification Polymerase Chain Reaction Polymorphism, Restriction Fragment Length Protozoan Proteins/biosynthesis/chemistry/*immunology *Protozoan Vaccines Restriction Mapping Sequence Homology, Amino Acid AD - Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8784778 ID - 176 ER - TY - JOUR AU - Leung, P. S. AU - Van de Water, J. AU - Coppel, R. L. AU - Nakanuma, Y. AU - Munoz, S. AU - Gershwin, M. E. PY - 1996 TI - Molecular aspects and the pathological basis of primary biliary cirrhosis SP - 119-28 N1 - Apr JF - J Autoimmun JO - Journal of autoimmunity VL - 9 IS - 2 SN - 0896-8411 (Print) N1 - Molecular aspects and the pathological basis of primary biliary cirrhosis N1 - 8738955 N1 - Ai 31585/ai/niaid Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review England N1 - eng KW - Animals Antibodies/immunology Epithelium/immunology Genes, Immunoglobulin Humans Liver Cirrhosis, Biliary/*immunology/pathology Mitochondria/immunology T-Lymphocytes/immunology N2 - Primary biliary cirrhosis (PBC) has been considered to be a 'model auto-immune disease' for more than two decades. However, the underlying pathophysiology of PBC and the relationship with the associated serological abnormalities have been hitherto elusive. Beginning in 1987 with the cloning and subsequent identification of the mitochondrial autoantigens of PBC, progress has come rapidly and we can now sketch several potential pathogenic pathways through which disease occurs. More than 90% of patients with PBC produce autoantibodies to mitochondria, and the antoantigens involved have been identified as related components of the 2-oxo-acid dehydrogenase complexes (OADC), including the E2 subunits of the pyruvate dehydrogenase complex (PDC-E2), the branched chain 2-oxo-acid dehdrogenase complex and 2-oxo-glutarate dehydrogenase complex, Protein X and E1 alpha. The cDNAs of each of the E2 subunits of OADC have been cloned and characterized. Moreover, the epitopes of the antimitochondrial antibodies (AMA) have been mapped at the highly conserved lipoyl domain E2 subunits. The use of recombinant peptides produced by these clones has greatly facilitated the detection of AMA. In addition, nucleotide sequence analysis of PDC-E2 specific human monoclonals and combinatorial Fabs strongly suggests that these autoantibodies are derived from clonal selection of a restricted set of somatically mutated immunoglobulin germline genes. Most interestingly, however, the use of monoclonal reagents to PDC-E2 has demonstrated that there is an increased expression of either PDC-E2, or a cross-reactive molecule, on the luminal surface of biliary epithelial cells in patients with PBC. These data provide a scenario to explain the tissue specific pathology associated with PBC and several interesting underlying pathophysiological mechanisms. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8738955 ID - 177 ER - TY - JOUR AU - Leung, P. S. AU - Cha, S. AU - Joplin, R. E. AU - Galperin, C. AU - Van de Water, J. AU - Ansari, A. A. AU - Coppel, R. L. AU - Schatz, P. J. AU - Cwirla, S. AU - Fabris, L. E. AU - Neuberger, J. M. AU - Gershwin, M. E. PY - 1996 TI - Inhibition of PDC-E2 human combinatorial autoantibodies by peptide mimotopes SP - 785-93 N1 - Dec JF - J Autoimmun JO - Journal of autoimmunity VL - 9 IS - 6 SN - 0896-8411 (Print) N1 - Inhibition of PDC-E2 human combinatorial autoantibodies by peptide mimotopes N1 - 9115581 N1 - Ai 31585/ai/niaid Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Amino Acid Sequence Animals Antibodies, Monoclonal *Autoantibodies Autoantigens/chemistry Biliary Tract/enzymology/immunology Epithelium/enzymology/immunology Humans Liver Cirrhosis, Biliary/enzymology/immunology Mice Molecular Mimicry Peptide Library Peptides/chemistry/immunology Pyruvate Dehydrogenase Complex/chemistry/*immunology Rabbits N2 - Immunohistochemical studies have shown that a unique immunoreactive molecule is present near the apical region of human biliary epithelial (BE) cells in patients with primary biliary cirrhosis (PBC). This can be visualized by confocal microscopy in PBC livers using a number of unique monoclonal antibodies to the E2 component of pyruvate dehydrogenase complex (PDC-E2), the autoantigen most commonly recognized by antimitochondrial antibodies (AMA). One such antibody, the murine mAb C355.1 was used to identify peptide mimotopes of PDC-E2 by screening a random dodecapeptide phage library ON 159.2 to identify the possible biochemical nature of this apical staining molecule. Out of 36 independent clones, 29 showed a common sequence and seven other sequences were singly represented. Three common amino acid motifs (SYP, TYVS and VRH) were found among these eight sequences. Similar to C355.1, the human combinatorial antibodies derived from a patient with PBC, SP1 and SP4, recognize the inner lipoyl domain of PDC-E2. However, when these antibodies are used to stain PBC BE cells, SP4 stains the apical region of PBC BE cells with high intensity whereas SP1 produces only cytoplasmic staining. Competitive inhibition of immunohistochemical staining using PDC-E2 specific human combinatorial antibodies SP1 and SP4 was performed using five of the above dodecapeptides. Interestingly, the peptides selected with C355.1 differentially inhibited the binding of SP1 and SP4 to PBC BE cells. Finally, rabbit sera raised against one such peptide (WMSYPDRTLRTS) stained BE cells from patients with PBC with a higher intensity than controls. Comparable data was obtained with immunoelectronmicroscopy. These data suggest that a molecular mimic of PDC-E2 is present at the external aspect of PBC BE cells. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis, School of Medicine 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9115581 ID - 168 ER - TY - JOUR AU - Galperin, C. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1996 TI - Use of recombinant proteins in the diagnosis of autoimmune connective tissue diseases SP - 337-47 N1 - Dec JF - Int Arch Allergy Immunol JO - International archives of allergy and immunology VL - 111 IS - 4 SN - 1018-2438 (Print) N1 - Use of recombinant proteins in the diagnosis of autoimmune connective tissue diseases N1 - 8957106 N1 - Journal Article Review Switzerland N1 - eng KW - Autoantibodies/analysis Autoantigens/*diagnostic use Autoimmune Diseases/*diagnosis Autoimmunity Connective Tissue Diseases/*diagnosis Humans Recombinant Proteins/*diagnostic use N2 - The use of recombinant proteins has been an invaluable tool for the study of the structure and function of cell components, and the analysis of autoimmune-mediated responses. Furthermore, it has also been instrumental in facilitating the development of reliable assays for the detection of a number of autoantibodies. This article discusses basic aspects of recombinant DNA technology, as well as its applications and limitations in the diagnosis of autoimmune connective tissue diseases such as systemic lupus erythematosus, Sjogren's syndrome, systemic scleroderma, mixed connective tissue disease, autoimmune inflammatory myopathies and primary vasculitides. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis, School of Medicine, 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8957106 ID - 169 ER - TY - JOUR AU - Dame, J. B. AU - Arnot, D. E. AU - Bourke, P. F. AU - Chakrabarti, D. AU - Christodoulou, Z. AU - Coppel, R. L. AU - Cowman, A. F. AU - Craig, A. G. AU - Fischer, K. AU - Foster, J. AU - Goodman, N. AU - Hinterberg, K. AU - Holder, A. A. AU - Holt, D. C. AU - Kemp, D. J. AU - Lanzer, M. AU - Lim, A. AU - Newbold, C. I. AU - Ravetch, J. V. AU - Reddy, G. R. AU - Rubio, J. AU - Schuster, S. M. AU - Su, X. Z. AU - Thompson, J. K. AU - Werner, E. B. AU - et al. PY - 1996 TI - Current status of the Plasmodium falciparum genome project SP - 1-12 N1 - Jul JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 79 IS - 1 SN - 0166-6851 (Print) N1 - Current status of the Plasmodium falciparum genome project N1 - 8844667 N1 - Journal Article Multicenter Study Research Support, Non-U.S. Gov't Review Netherlands N1 - eng KW - Animals Cell Nucleus/genetics Chromosome Mapping DNA, Complementary/genetics Gene Expression Genes, Protozoan *Genome, Protozoan Molecular Sequence Data Organizations Plasmodium falciparum/*genetics Sequence Analysis, DNA N2 - The Plasmodium falciparum Genome Project is a collaborative effort by many laboratories that will provide detailed molecular information about the parasite, which may be used for developing practical control measures. Initial goals are to prepare an electronically indexed clone bank containing partially sequenced clones representing up to 80% of the parasite's genes and to prepare an ordered set of overlapping clones spanning each of the parasite's 14 chromosomes. Currently, clones of genomic DNA, prepared as yeast artificial chromosomes, are arranged into contigs covering approximately 70% of the genome of parasite clone 3D7, gene sequence tags are available from more than contigs covering approximately 70% of the genome of parasite clone 3D7, gene sequence tags are available from more than 20% of the parasite's genes, and approximately 5% of the parasite's genes are tentatively identified from similarity searches of entries in the international sequence databases. A total of > 0.5 Mb of P. falciparum sequence tag data is available. The gene sequence tags are presently being used to complete YAC contig assembly and localize the cloned genes to positions on the physical map in preparation for sequencing the genome. Routes of access to project information and services are described. AD - University of Florida, Gainesville, 32611, USA. dame@icbr.ifas.ufl.edu UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8844667 ID - 174 ER - TY - JOUR AU - Cooke, B. M. AU - Rogerson, S. J. AU - Brown, G. V. AU - Coppel, R. L. PY - 1996 TI - Adhesion of malaria-infected red blood cells to chondroitin sulfate A under flow conditions SP - 4040-4 N1 - Nov 15 JF - Blood JO - Blood VL - 88 IS - 10 SN - 0006-4971 (Print) N1 - Adhesion of malaria-infected red blood cells to chondroitin sulfate A under flow conditions N1 - 8916971 N1 - Dk32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Cell Adhesion Chondroitin Sulfates/*metabolism Dose-Response Relationship, Drug Erythrocytes/*parasitology/pathology Hemorheology Humans Malaria/*blood/parasitology Stress, Mechanical N2 - Adhesion of parasitized red blood cells (PRBCs) to microvascular endothelial cells (ECs) is a distinctive feature of Plasmodium falciparum malaria and is a central event in the development of life-threatening complications such as cerebral malaria. PRBCs adhere to several EC-expressed molecules in vitro, but the relative importance of these interactions in vivo remains unclear. Chondroitin sulfate A (CSA) is the most recent EC surface-associated molecule to be implicated in the adhesive process. Accordingly, we have studied adhesion of PRBCs to CSA in vitro using a parallel-plate flow chamber. Under controlled flow conditions, PRBCs adhered to CSA in a concentration-dependent manner at wall-shear stresses up to 0.2 Pa, a value that is within the physiological range for venules. Once adhered, PRBCs remained stationary (rather than rolling) and continued to remain stationary even when the wall-shear stress was raised to supravenular levels. The adhesive interaction was strong and a proportion of adherent PRBCs could withstand detachment at stresses up to 2.5 Pa. Soluble CSA at pharmacological concentrations prevented adhesion of flowing PRBCs in a concentration-dependent manner but failed to reverse established adhesion. Adhesion of PRBCs to CSA could contribute to the pathogenesis of malaria, and soluble CSA may have a useful therapeutic effect. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8916971 ID - 170 ER - TY - JOUR AU - Cha, S. AU - Leung, P. S. AU - Van de Water, J. AU - Tsuneyama, K. AU - Joplin, R. E. AU - Ansari, A. A. AU - Nakanuma, Y. AU - Schatz, P. J. AU - Cwirla, S. AU - Fabris, L. E. AU - Neuberger, J. M. AU - Gershwin, M. E. AU - Coppel, R. L. PY - 1996 TI - Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2) SP - 10949-54 N1 - Oct 1 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 93 IS - 20 SN - 0027-8424 (Print) N1 - Random phage mimotopes recognized by monoclonal antibodies against the pyruvate dehydrogenase complex-E2 (PDC-E2) N1 - 8855289 N1 - Ai 31585/ai/niaid Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Amino Acid Sequence Animals Antibodies, Monoclonal/*immunology Autoantigens/*chemistry/immunology Bile Ducts/immunology Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/immunology Epitope Mapping Fluorescent Antibody Technique, Indirect Humans Liver Cirrhosis, Biliary/*immunology Molecular Mimicry Peptide Library Peptides/immunology Pyruvate Dehydrogenase Complex/chemistry/*immunology Rabbits N2 - Dihydrolipoamide acetyltransferase, the E2 component of the pyruvate dehydrogenase complex (PDC-E2), is the autoantigen most commonly recognized by autoantibodies in primary biliary cirrhosis (PBC). We identified a peptide mimotope(s) of PDC-E2 by screening a phage-epitope library expressing random dodecapeptides in the pIII coat protein of fd phage using C355.1, a murine monoclonal antibody (mAb) that recognizes a conformation-dependent epitope in the inner lipoyl domain of PDC-E2 and uniquely stains the apical region of bile duct epithelium (BDE) only in patients with PBC. Eight different sequences were identified in 36 phage clones. WMSYPDRTLRTS was present in 29 clones; WESYPFRVGTSL, APKTYVSVSGMV, LTYVSLQGRQGH, LDYVPLKHRHRH, AALWGVKVRHVS, KVLNRIMAGVRH and GNVALVSSRVNA were singly represented. Three common amino acid motifs (W-SYP, TYVS, and VRH) were shared among all peptide sequences. Competitive inhibition of the immunohistochemical staining of PBC BDE was performed by incubating the peptides WMSYPDRTLRTS, WESYPDRTLRTS, APKTYVSVSGMV, and AALWGVKVRHVS with either C355.1 or a second PDC-E2-specific mAb, C150.1. Both mAbs were originally generated to PDC-E2 but map to distinct regions of PDC-E2. Two of the peptides, although selected by reaction with C355.1, strongly inhibited the staining of BDE by C150.1, whereas the peptide APKTYVSVSGMV consistently inhibited the staining of C355.1 on biliary duct epithelium more strongly than the typical mitochondrial staining of hepatocytes. Rabbit sera raised against the peptide WMSYPDRTLRTS stained BDE of livers and isolated bile duct epithelial cells of PBC patients more intensively than controls. The rabbit sera stained all size ducts in normals, but only small/medium-sized ductules in PBC livers. These studies provide evidence that the antigen present in BDE is a molecular mimic of PDC-E2, and not PDC-E2 itself. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8855289 ID - 173 ER - TY - JOUR AU - Billman-Jacobe, H. AU - Sloan, J. AU - Coppel, R. L. PY - 1996 TI - Analysis of isoniazid-resistant transposon mutants of Mycobacterium smegmatis SP - 47-52 N1 - Oct 15 JF - FEMS Microbiol Lett JO - FEMS microbiology letters VL - 144 IS - 1 SN - 0378-1097 (Print) N1 - Analysis of isoniazid-resistant transposon mutants of Mycobacterium smegmatis N1 - 8870251 N1 - Comparative Study Journal Article Netherlands N1 - eng KW - *Bacterial Proteins Chromosome Mapping DNA Transposable Elements Drug Resistance, Microbial/genetics Genetic Markers Isoniazid/*pharmacology Mutagenesis, Insertional *Mutation Mycobacterium/*drug effects/*genetics Peroxidase/analysis Peroxidases/*genetics Polymerase Chain Reaction Sequence Analysis, DNA N2 - The emergence of multidrug-resistant tuberculosis has renewed interest in the study of drug resistance in mycobacteria with the objective of improved chemotherapy. The genetic basis of isoniazid resistance in a model mycobacterium was studied. Eleven isoniazid-resistant mutants of Mycobacterium smegmatis were created using transposon mutagenesis. Genetic and enzymatic characterisation of the mutants showed that katG, encoding T-catalase, was inactivated. The nucleotide sequence of M. smegmatis katG was determined and the mutation sites mapped demonstrating that both the amino and carboxyl halves of T-catalase are important for enzymatic activity. AD - Department of Microbiology, Monash University, Clayton, Vic., Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8870251 ID - 172 ER - TY - JOUR AU - Waterkeyn, J. G. AU - Lightowlers, M. W. AU - Coppel, R. AU - Cowman, A. F. PY - 1995 TI - Characterization of the gene family encoding a host-protective antigen of the tapeworm Taenia ovis SP - 123-31 N1 - Jul JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 73 IS - 1-2 SN - 0166-6851 (Print) N1 - Characterization of the gene family encoding a host-protective antigen of the tapeworm Taenia ovis N1 - 8577320 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigenic Variation Antigens, Helminth/*genetics Base Sequence DNA, Complementary/genetics DNA, Helminth/genetics *Genes, Helminth Molecular Sequence Data *Multigene Family RNA, Helminth/genetics RNA, Messenger/genetics Sequence Homology, Nucleic Acid Taenia/*genetics/*immunology N2 - Genomic structure has been determined for a gene encoding a host-protective antigen of the parasitic platyhelminth Taenia ovis. An incomplete cDNA, known as 45W, encodes the protective antigen. Southern hybridisation experiments using 45W cDNA as a probe, revealed that the 45W gene was a member of a multigene family. Differential Southern hybridisation and rapid amplification of cDNA end (RACE) experiments were used to characterise the related genes, allowing the full-length coding region of the 45W encoded antigen to be determined. The gene family comprises a minimum of four members per haploid genome with each member showing varying degrees of 5' and 3' homology with respect to the 45W cDNA. A close homologue of the 45W gene, designated 45S, differed from 45W at 11 of 985 nt comprising the full-length mRNA. Sequencing of several independent RACE products for both 45W and 45S identified a cDNA which may be a product of homologous recombination between these genes, suggesting that the two genes may be alleles. Homologous recombination in genes which encode a host protective antigen such as 45W would provide a mechanism by which antigenic variants could arise. AD - University of Melbourne, Veterinary Clinical Centre, Werribee, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8577320 ID - 184 ER - TY - JOUR AU - Van de Water, J. AU - Ansari, A. AU - Prindiville, T. AU - Coppel, R. L. AU - Ricalton, N. AU - Kotzin, B. L. AU - Liu, S. AU - Roche, T. E. AU - Krams, S. M. AU - Munoz, S. AU - Gershwin, M. E. PY - 1995 TI - Heterogeneity of autoreactive T cell clones specific for the E2 component of the pyruvate dehydrogenase complex in primary biliary cirrhosis SP - 723-33 N1 - Feb 1 JF - J Exp Med JO - The Journal of experimental medicine VL - 181 IS - 2 SN - 0022-1007 (Print) N1 - Heterogeneity of autoreactive T cell clones specific for the E2 component of the pyruvate dehydrogenase complex in primary biliary cirrhosis N1 - 7836925 N1 - 1-dk-6-2274/dk/niddk Ar-37070/ar/niams Dk-39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged Autoimmunity Clone Cells Female Humans Liver/immunology/pathology Liver Cirrhosis, Biliary/enzymology/*immunology Middle Aged Mitochondria, Liver/immunology Monocytes/immunology Phenotype Pyruvate Dehydrogenase Complex/*immunology Receptors, Antigen, T-Cell/immunology T-Lymphocytes/*immunology N2 - The extraordinary specificity of bile duct destruction in primary biliary cirrhosis (PBC) and the presence of T cell infiltrates in the portal tracts have suggested that biliary epithelial cells are the targets of an autoimmune response. The immunodominant antimitochondrial response in patients with PBC is directed against the E2 component of pyruvate dehydrogenase (PDC-E2). Hitherto, there have only been limited reports on the characterization and V beta usage of PDC-E2-specific cloned T cell lines. In this study, we examined peripheral blood mononuclear cells (PBMC) for their reactivity to the entire PDC complex as well as to the E1- and E2-specific components. We also examined the phenotype, lymphokine profile, and V beta usage of PDC-specific T cell clones isolated from cellular infiltrates from the livers of PBC patients. We report that PBMC from 16/19 patients with PBC, but not 12 control patients, respond to the PDC-E2 subunit. Interestingly, this response was directed to the inner and/or the outer lipoyl domains, despite the serologic observation that the autoantibody response is directed predominantly to the inner lipoyl domain. Additionally, lymphokine analysis of interleukin (IL) 2/IL-4/interferon gamma production from individual liver-derived autoantigen-specific T cell clones suggests that both T helper cell Th1- and Th2-like clones are present in the liver. Moreover, there was considerable heterogeneity in the T cell receptor for antigen (TCR) V beta usage of these antigen-specific autoreactive T cell clones. This is in contrast to murine studies in which animals are induced to develop autoimmunity by specific immunization and have an extremely limited T cell V beta repertoire. Thus, our data suggest that in human organ-specific autoimmune diseases, such as PBC, the TCR V beta repertoire is heterogenous. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7836925 ID - 188 ER - TY - JOUR AU - Tsuneyama, K. AU - Van De Water, J. AU - Van Thiel, D. AU - Coppel, R. AU - Ruebner, B. AU - Nakanuma, Y. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1995 TI - Abnormal expression of PDC-E2 on the apical surface of biliary epithelial cells in patients with antimitochondrial antibody-negative primary biliary cirrhosis SP - 1440-6 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 22 IS - 5 SN - 0270-9139 (Print) N1 - Abnormal expression of PDC-E2 on the apical surface of biliary epithelial cells in patients with antimitochondrial antibody-negative primary biliary cirrhosis N1 - 7590661 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged Aged, 80 and over Animals Autoantibodies/analysis Autoantigens/analysis Bile Ducts, Intrahepatic/enzymology/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/enzymology/immunology Female Histocompatibility Antigens Class II/analysis Humans Liver Cirrhosis, Biliary/enzymology/*immunology Male Mice Microscopy, Confocal Middle Aged Mitochondria/immunology Pyruvate Dehydrogenase Complex/*analysis Rabbits Rheology N2 - The presence of antimitochondrial antibodies (AMA) is a major criterion for the diagnosis of primary biliary cirrhosis (PBC). Although it is not clear that AMA are involved in the pathogenesis of the disease, the study of these autoantibodies has enabled much information to be accumulated about the specificity of this response. The autoantigens have been identified as components of a functionally related enzyme family, the 2-oxo-acid-dehydrogenase complex. Within this complex, pyruvate dehydrogenase E2 subunit (PDC-E2) has been determined to be the immunodominant autoantigen. Using a panel of mouse monoclonal antibodies and human combinatorial autoantibodies, it has been demonstrated that patients with PBC, but not controls, have an abnormal expression of either PDC-E2 or a cross-reacting molecule in the apical region of biliary epithelium. Others have shown a similar reaction using rabbit sera directed to PDC-E2. Our previous studies have concentrated on AMA-positive patients. In this study, the presence of PDC-E2, class II, immunoglobulin (Ig) A, and B7/BB1 in the bile duct epithelial cells of AMA-positive as well as AMA-negative patients is addressed. Most patients with AMA-negative PBC (seven of nine) react in a fashion similar to AMA-positive patients with intense staining of the apical region of the bile duct epithelial cells of "PDC-E2," increased IgA expression, and little major histocompatibility complex (MHC) class II staining in the early-stage patients. Interestingly, the two AMA-negative patients that did not express PDC-E2 on the apical side of their biliary epithelium had anticentromere antibodies and Sjogren's syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Division of Rheumatology, Allergy and Clinical Immunology University of California Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7590661 ID - 179 ER - TY - JOUR AU - Tsuneyama, K. AU - Van de Water, J. AU - Leung, P. S. AU - Cha, S. AU - Nakanuma, Y. AU - Kaplan, M. AU - De Lellis, R. AU - Coppel, R. AU - Ansari, A. AU - Gershwin, M. E. PY - 1995 TI - Abnormal expression of the E2 component of the pyruvate dehydrogenase complex on the luminal surface of biliary epithelium occurs before major histocompatibility complex class II and BB1/B7 expression SP - 1031-7 N1 - Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 21 IS - 4 SN - 0270-9139 (Print) N1 - Abnormal expression of the E2 component of the pyruvate dehydrogenase complex on the luminal surface of biliary epithelium occurs before major histocompatibility complex class II and BB1/B7 expression N1 - 7535733 N1 - 1-dk-6-22740/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Animals Antigens, CD80/*analysis Bile Ducts/enzymology/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Female HLA-DR Antigens/*analysis Humans Immunoglobulin A/analysis Liver Cirrhosis, Biliary/enzymology/*immunology Mice Middle Aged Pyruvate Dehydrogenase Complex/*analysis/immunology N2 - Primary biliary cirrhosis (PBC) is a chronic autoimmune liver disease characterized histologically by nonsuppurative destructive cholangitis. Sera from patients with PBC react with a series of intramitochondrial enzymes with the immunodominant response directed against the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Recently, using tissue sections of late-stage PBC, we showed that there is increased expression in biliary epithelial cells of patients with PDC-E2 or a molecule cross-reactive with PDC-E2. Previous work has shown that biliary epithelial cells of patients with PBC express an increased amount of class II. To address the sequence of events in the evolution of PBC, we have focused our attention in this study on early biliary epithelial lesions. In particular, we have studied the liver of 22 female patients with PBC that was diagnosed as either stage I or stage II using both a mouse monoclonal antibody that has reactivity similar to human autoantibodies as well as a human Fab combinatorial prepared from the lymph node of a PBC patient. Tissues were simultaneously stained using antibodies to PDC-E2, class II, and BB1/B7. As a positive control, tissues from late-stage PBC were studied concurrently. By determining the order of expression among the three molecules, PDC-E2, class II, and BB1/B7, we report that the expression of PDC-E2 or a PDC-E2-like molecule on biliary duct epithelium of patients with PBC precedes the expression of BB1/B7 and major histocompatibility complex (MHC) class II molecules. The alteration of an autoantigen in biliary duct epithelium may be the earliest lesion in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7535733 ID - 186 ER - TY - JOUR AU - Sturchler, D. AU - Berger, R. AU - Rudin, C. AU - Just, M. AU - Saul, A. AU - Rzepczyk, C. AU - Brown, G. AU - Anders, R. AU - Coppel, R. AU - Woodrow, G. AU - et al. PY - 1995 TI - Safety, immunogenicity, and pilot efficacy of Plasmodium falciparum sporozoite and asexual blood-stage combination vaccine in Swiss adults SP - 423-31 N1 - Oct JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 53 IS - 4 SN - 0002-9637 (Print) N1 - Safety, immunogenicity, and pilot efficacy of Plasmodium falciparum sporozoite and asexual blood-stage combination vaccine in Swiss adults N1 - 7485698 N1 - Clinical Trial Clinical Trial, Phase I Journal Article Randomized Controlled Trial Research Support, Non-U.S. Gov't United states N1 - eng KW - Adolescent Adult Amino Acid Sequence Animals Antibodies, Protozoan/biosynthesis Antigens, Protozoan/chemistry/genetics/immunology Antigens, Surface/chemistry/genetics/immunology Enzyme-Linked Immunosorbent Assay Female Fluorescent Antibody Technique, Indirect Humans Malaria Vaccines/adverse effects/immunology/standards Malaria, Falciparum/prevention & control Male Merozoite Surface Protein 1 Middle Aged Molecular Sequence Data Pilot Projects Plasmodium falciparum/*immunology Protein Precursors/chemistry/genetics/immunology Protozoan Proteins Protozoan Vaccines/adverse effects/*immunology/standards Single-Blind Method Vaccines, Synthetic N2 - This study was part of a larger program to develop a vaccine effective against Plasmodium falciparum infection caused by sporozoites and clinical malaria caused by asexual blood stages. In a phase 1 study of safety and immunogenicity, two recombinant proteins (Ro 46-2717, a circumsporozoite [CS] protein) construct with a molecular mass of 35 kD, and Ro 46-2924, a merozoite surface antigen [MSA-2] construct with a molecular mass of 25 kD) adsorbed onto alum were injected in two low (20 micrograms) or two high (100 micrograms) doses in the right and left deltoid muscles of 33 healthy Swiss volunteers; six other volunteers received a placebo (alum alone). Twenty-six participants reported 51 immunization-related adverse events, mainly pain at the injection site. Mean antibody titers to CS protein and MSA-2 in an indirect immunofluorescence assay peaked four weeks after the second immunization without evidence of boosting (i.e., sharp increase in titer). By that time, 56% and 31% of the vaccinees seroconverted to CS protein and MSA-2, respectively, with the increase in MSA-2 titer being weaker than that for the CS protein. After a third immunization, five vaccinees volunteered to be challenged by three or four infective bites of Anopheles stephensi. Prepatent and incubation periods in all five were comparable with unvaccinated historic controls challenged under similar conditions, and all had symptoms of clinical falciparum malaria. We conclude that the vaccine components were safe and immunogenic but there was no evidence that this immunization regimen with the CS protein plus MSA-2 component was able to prevent infection.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Tropical Medicine Unit, F. Hoffmann-La Roche & Co., Ltd., Basel, Switzerland. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7485698 ID - 181 ER - TY - JOUR AU - Magowan, C. AU - Coppel, R. L. AU - Lau, A. O. AU - Moronne, M. M. AU - Tchernia, G. AU - Mohandas, N. PY - 1995 TI - Role of the Plasmodium falciparum mature-parasite-infected erythrocyte surface antigen (MESA/PfEMP-2) in malarial infection of erythrocytes SP - 3196-204 N1 - Oct 15 JF - Blood JO - Blood VL - 86 IS - 8 SN - 0006-4971 (Print) N1 - Role of the Plasmodium falciparum mature-parasite-infected erythrocyte surface antigen (MESA/PfEMP-2) in malarial infection of erythrocytes N1 - 7579415 N1 - Dk 32094/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Antigens, Protozoan/*physiology Cell Adhesion *Cytoskeletal Proteins Erythrocytes/*parasitology Host-Parasite Relations Malaria, Falciparum/*blood Melanoma/pathology Membrane Proteins/deficiency/*metabolism Microscopy, Confocal *Neuropeptides Peptides/physiology Plasmodium falciparum/immunology/*physiology Protein Binding Protozoan Proteins/*physiology Rabbits Tumor Cells, Cultured N2 - During intraerythrocytic growth of Plasmodium falciparum, several parasite proteins are transported from the parasite to the erythrocyte membrane, where they bind to membrane skeletal proteins. Mature-parasite-infected erythrocyte surface antigen (MESA) has previously been shown to associate with host erythrocyte membrane skeletal protein 4.1. Using a spontaneous mutant of P falciparum that has lost the ability to synthesize MESA and 4.1-deficient erythrocytes, we examined growth of MESA(+) and MESA(-) parasites in normal and 4.1-deficient erythrocytes. Viability of MESA(+) parasites was reduced in 4.1-deficient erythrocytes as compared with that for normal erythrocytes, but MESA(-) parasites grew equally well in 4.1-deficient and normal erythrocytes. Cytoadherence of MESA(+)- and MESA (-)-parasitized normal and 4.1-deficient erythrocytes to C32 melanoma cells was similar, indicating that neither protein 4.1 nor MESA plays a major role in cytoadherence of infected erythrocytes. Localization of MESA in normal and 4.1-deficient erythrocytes was examined by confocal microscopy. MESA was diffusely distributed in the cytosol of 4.1-deficient erythrocytes but was membrane-associated in normal erythrocytes. These findings suggest that MESA binding to protein 4.1 plays a major role in intraerythrocytic parasite viability. AD - Life Sciences Division, Lawrence Berkeley Laboratory, Berkeley, CA, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7579415 ID - 180 ER - TY - JOUR AU - Leung, P. S. AU - Chuang, D. T. AU - Wynn, R. M. AU - Cha, S. AU - Danner, D. J. AU - Ansari, A. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1995 TI - Autoantibodies to BCOADC-E2 in patients with primary biliary cirrhosis recognize a conformational epitope SP - 505-13 N1 - Aug JF - Hepatology JO - Hepatology (Baltimore, Md VL - 22 IS - 2 SN - 0270-9139 (Print) N1 - Autoantibodies to BCOADC-E2 in patients with primary biliary cirrhosis recognize a conformational epitope N1 - 7543435 N1 - 1-dk-6-2274/dk/niddk Ai 31585/ai/niaid Dk 39588/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Amino Acid Sequence Antibody Specificity Autoantibodies/*blood Autoantigens/chemistry/immunology Binding Sites Enzyme-Linked Immunosorbent Assay Epitopes/*chemistry/immunology Humans Immunosorbent Techniques Ketone Oxidoreductases/*immunology Liver Cirrhosis, Biliary/*immunology Molecular Sequence Data Multienzyme Complexes/*immunology Protein Conformation Recombinant Proteins/analysis/chemistry/immunology Sequence Analysis Sequence Homology Thioctic Acid/analysis/metabolism N2 - Primary biliary cirrhosis (PBC) is an autoimmune disease of liver associated with a unique serologic response to mitochondrial autoantigens. Many of the autoantigens recognized by autoantibodies in PBC are members of the 2-oxo-acid dehydrogenase complex. The two major autoantigens are the E2 component of the pyruvate dehydrogenase complex (PDC-E2) and the E2 component of the branched chain 2-oxo-acid dehydrogenase complex (BCOADC-E2). The autoantibody response to PDC-E2 has been mapped to one immunodominant epitope, which consists of both linear and conformational components. The presence of a single immunodominant epitope in PDC-E2 is unusual when contrasted to the immune response to autoantigens in other human autoimmune diseases. We have mapped the epitope recognized by antimitochondrial autoantibodies (AMA) specific to BCOADC-E2 in patients with PBC by taking advantage of the full-length bovine BCOADC-E2 complementary DNA (cDNA) and a series of expression clones spanning the entire molecule. Reactivity to the various expression clones was studied by immunoblotting, enzyme-linked immunosorbent assay (ELISA), as well as selective absorption of patient sera by expressed protein fragments. Autoantibodies to BCOADC-E2 map within peptides spanning amino acid residues 1 to 227 of the mature protein; our data demonstrate that the epitope is dependent on conformation and includes the lipoic acid binding region. However, only the full-length clone (amino acid residue 1 to 421) is sufficient to remove all detectable BCOADC-E2 reactivity. Moreover, the absence of lipoic acid on the recombinant polypeptides used in this study indicates that antibody binding to BCOADC-E2 is not dependent on the presence of lipoic acid. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616, USA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7543435 ID - 183 ER - TY - JOUR AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1995 TI - Primary biliary cirrhosis: the molecule and the mimic SP - 17-49 N1 - Apr JF - Immunol Rev JO - Immunological reviews VL - 144 SN - 0105-2896 (Print) N1 - Primary biliary cirrhosis: the molecule and the mimic N1 - 7590813 N1 - 1-dk-6-2274/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review Denmark N1 - eng KW - Amino Acid Sequence Autoimmune Diseases/*immunology Humans Liver Cirrhosis, Biliary/*immunology Molecular Sequence Data N2 - Our understanding of the immunobiology of PBC has dramatically changed with the application of molecular biology to clinical medicine. Because of the molecular characterization and identification of the mitochondrial autoantigens, it is now possible to define explicitly mitochondrial autoantigens and examine recognition sites at the primary sequence level. In addition, the expression of cloned antigens has facilitated the development of more reliable assays for mitochondrial autoantibodies. The use of cloned recombinant antigens should, one day, replace the traditional AMA immunofluorescence for diagnostic assays. Possible genetic and environmental factors associated with risk for PBC can also be investigated. It is now also possible to begin the task to defining the role of T cells in the immunopathology of PBC and exploring the issue of whether specific immunotherapy is feasible. There is increasing evidence that PDC-E2 or a similar molecule is located on the cell membrane of biliary epithelial cells. The mechanism for this expression remains to be studied. The explosion of data in PBC is an example of the application of new techniques to investigate old problems. This has occurred because of networking between laboratories in many countries and the generous exchange of sera and donation of livers removed at transplantation. Unfortunately, there is no animal model for PBC; if an animal model was found it would have major importance. Finally, we emphasize the need to study patients early in the course of disease in order to define the events that initiate pathology. AD - Monash University, Department of Microbiology, Clayton, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7590813 ID - 185 ER - TY - JOUR AU - Cooke, B. M. AU - Coppel, R. L. PY - 1995 TI - Cytoadhesion and falciparum malaria: going with the flow SP - 282-7 N1 - Aug JF - Parasitol Today JO - Parasitology today (Personal ed VL - 11 IS - 8 SN - 0169-4758 (Print) N1 - Cytoadhesion and falciparum malaria: going with the flow N1 - 15275324 N1 - Journal Article England N1 - eng N2 - Sequestration of parasitized red blood cells in the cerebral vasculature is the predisposing event to the development of cerebral malaria during infection with Plasmodium falciparum. The adhesive interaction between these cells and receptors on the endothelial cell (cytoadhesion) occurs in the dynamic environment of the microcirculation, but most studies have neglected this factor and have concentrated on measuring adhesion in static (no flow) assays. Such studies ignore the markedly different rheological properties of parasitized red blood cells that become apparent when adhesion is examined under dynamic, flow conditions that resemble those of the circulation in vivo. Here, Brian Cooke and Ross Coppel review a number of novel aspects of cytoadhesion that have been identified using flow-based assays, and discuss their relevance to the pathophysiology, investigation and clinical management of falciparum malaria. AD - Department of Microbiology, Monash University, Clayton, Victoria, Australia. brian.cooke@med.monash.edu.au UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15275324 ID - 182 ER - TY - JOUR AU - Bennett, B. J. AU - Thompson, J. AU - Coppel, R. L. PY - 1995 TI - Identification of Plasmodium falciparum histone 2B and histone 3 genes SP - 231-3 N1 - Mar JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 70 IS - 1-2 SN - 0166-6851 (Print) N1 - Identification of Plasmodium falciparum histone 2B and histone 3 genes N1 - 7637710 N1 - Dk-32094-10/dk/niddk Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Amino Acid Sequence Animals DNA, Protozoan/genetics Genes, Protozoan/*genetics Histones/*genetics Invertebrates/genetics Molecular Sequence Data Plasmodium falciparum/*genetics Protozoan Proteins/*genetics Sequence Alignment Sequence Homology, Amino Acid Species Specificity AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7637710 ID - 187 ER - TY - JOUR AU - Tsuneyama, K. AU - Van de Water, J. AU - Nakanuma, Y. AU - Cha, S. AU - Ansari, A. AU - Coppel, R. AU - Gershwin, M. E. PY - 1994 TI - Human combinatorial autoantibodies and mouse monoclonal antibodies to PDC-E2 produce abnormal apical staining of salivary glands in patients with coexistent primary biliary cirrhosis and Sjogren's syndrome SP - 893-8 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 20 IS - 4 Pt 1 SN - 0270-9139 (Print) N1 - Human combinatorial autoantibodies and mouse monoclonal antibodies to PDC-E2 produce abnormal apical staining of salivary glands in patients with coexistent primary biliary cirrhosis and Sjogren's syndrome N1 - 7927231 N1 - 39588/phs Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adult Aged Animals Antibodies, Monoclonal/*immunology Antigen-Antibody Reactions Autoantibodies/*immunology Autoantigens/immunology/metabolism Bile Ducts/enzymology/immunology Cross Reactions Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/enzymology/immunology Female Humans Immunohistochemistry Liver Cirrhosis, Biliary/complications/enzymology/*immunology Male Mice Middle Aged Pyruvate Dehydrogenase Complex/immunology/*metabolism Salivary Glands/enzymology/*immunology Sjogren's Syndrome/complications/enzymology/*immunology N2 - An increase in the incidence of Sjogren's syndrome in patients with primary biliary cirrhosis has been noted. Indeed, primary biliary cirrhosis has been described as a ductal disease with involvement not only of the biliary tract but of epithelial ductal cells in other organs. We have previously reported the development of a panel of mouse monoclonal antibodies directed at PDC-E2, the major autoantigen of primary biliary cirrhosis. One such antibody, C355.1, but none of the other monoclonal antibodies, reacted not only with mitochondria but also with the apical region of biliary epithelium of patients with primary biliary cirrhosis but not in similar specimens from patients with other liver disease or normal human liver. In addition, we have reported the development of human combinatorial antibodies specific for PDC-E2; these reagents also reacted uniquely with the biliary epithelium of patients with primary biliary cirrhosis. In this paper, we have performed a similar study and have compared the staining of monoclonal antibody C355.1 and a human combinatorial antibody, SP4, with control monoclonal antibodies with respect to their reactivity of salivary glands in 9 patients with primary biliary cirrhosis associated with Sjogren's syndrome, 11 patients with Sjogren's syndrome alone and 7 control patients. Interestingly, the apical region of the salivary gland epithelial cells of approximately 50% of patients with coexisting primary biliary cirrhosis and Sjogren's syndrome had a staining pattern similar to that seen in primary biliary cirrhosis biliary epithelium. In contrast, we did not observe this reactivity in any patient with Sjogren's syndrome alone or in any control patient.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California--Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7927231 ID - 192 ER - TY - JOUR AU - Reeder, J. C. AU - Rogerson, S. J. AU - al-Yaman, F. AU - Anders, R. F. AU - Coppel, R. L. AU - Novakovic, S. AU - Alpers, M. P. AU - Brown, G. V. PY - 1994 TI - Diversity of agglutinating phenotype, cytoadherence, and rosette-forming characteristics of Plasmodium falciparum isolates from Papua New Guinean children SP - 45-55 N1 - Jul JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 51 IS - 1 SN - 0002-9637 (Print) N1 - Diversity of agglutinating phenotype, cytoadherence, and rosette-forming characteristics of Plasmodium falciparum isolates from Papua New Guinean children N1 - 8059915 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Agglutination Tests Animals Antigenic Variation Antigens, Protozoan/immunology Antigens, Surface/immunology Cell Adhesion Cell Line Child Child, Preschool Cryopreservation Female Humans Immune Sera/immunology Infant Malaria, Falciparum/*parasitology Male Melanoma, Amelanotic Papua New Guinea Phenotype Plasmodium falciparum/classification/*immunology Rosette Formation Tumor Cells, Cultured N2 - The relationship between antigenic variation, cytoadherence, rosette formation, and the pathogenesis of malaria has led to great interest in the diversity of these properties in Plasmodium falciparum isolates from different communities. In this study, we extend previous investigations by delineating the spectrum of agglutinating phenotypes, adherence to C32 melanoma cells, human umbilical vein endothelial cells (HUVEC), CD36, and intracellular adhesion molecule-1 (ICAM-1), and rosette-forming ability of a group of 20 P. falciparum isolates from Papua New Guinean children. Agglutination phenotypes were determined by using both the children's convalescent serum and a panel of adult immune sera. The wide range of variant antigenic types in the community was demonstrated by the failure of the agglutination assays to identify any two isolates with the same agglutinating phenotype in this, the largest study of its kind. Comparison of agglutination profiles from fresh and cryopreserved isolates demonstrated the general acceptability of cryopreservation before testing, but cautioned that some isolates may undergo selection and phenotypic change during the process. Nineteen isolates were able to bind to at least one of the four ligands studied and showed marked variation in both avidity and specificity of binding. The purified proteins ICAM-1 and CD36 proved to be the most useful assay ligands for investigating field isolates, with 18 isolates binding to at least one protein and 14 to both. No correlation was found between the binding of isolates to any two ligands nor between the binding of a standardized inoculum and the level of the patient's presenting parasitemia. All isolates from the study group were found to form rosettes (at a mean rate of 14.6% of cultured trophozoites involved in rosettes). A lack of correlation between rosette formation and CD36 binding suggests that the previously reported role of CD36 as a rosette formation receptor may not be important for isolates from Papua New Guinea. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8059915 ID - 195 ER - TY - JOUR AU - McColl, D. J. AU - Silva, A. AU - Foley, M. AU - Kun, J. F. AU - Favaloro, J. M. AU - Thompson, J. K. AU - Marshall, V. M. AU - Coppel, R. L. AU - Kemp, D. J. AU - Anders, R. F. PY - 1994 TI - Molecular variation in a novel polymorphic antigen associated with Plasmodium falciparum merozoites SP - 53-67 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 68 IS - 1 SN - 0166-6851 (Print) N1 - Molecular variation in a novel polymorphic antigen associated with Plasmodium falciparum merozoites N1 - 7891748 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan *Antigenic Variation Antigens, Protozoan/chemistry/*genetics Base Sequence Cloning, Molecular DNA, Complementary/genetics DNA, Protozoan/genetics Escherichia coli/genetics Genes, Protozoan Humans Malaria, Falciparum/immunology Molecular Sequence Data Molecular Weight Plasmodium falciparum/*genetics/growth & development/*immunology Polymorphism, Genetic Protozoan Proteins/chemistry/genetics/immunology RNA, Messenger/genetics RNA, Protozoan/genetics Repetitive Sequences, Nucleic Acid N2 - A cDNA clone encoding part of a novel polymorphic merozoite antigen from Plasmodium falciparum was isolated by screening a cDNA library with human immune serum from Papua New Guinea. Immunofluorescence microscopy and immunoblotting with affinity-purified antibodies recognized a highly polymorphic antigen, Ag956, present in schizonts and merozoites. Biosynthetic labeling and immunoprecipitation experiments demonstrated that Ag956 is proteolytically cleaved during merozoite maturation. The complete genomic sequence of Ag956 from the D10 clone of P. falciparum isolate FC27 encodes a secreted protein of calculated molecular mass 43,243 that is very hydrophilic and contains a region of unusual heptad repeats of the general structure AXXAXXX. This antigen has been named the secreted polymorphic antigen associated with merozoites (SPAM). The sequence of a second SPAM allele from the 3D7 clone of isolate NF54 reveals that the alanine heptad repeats and the hydrophilic C-terminal half of the protein are conserved. Variation among SPAM alleles is the result of deletions and amino acid substitutions in non-repetitive sequences within and flanking the alanine heptad-repeat domain. Heptad repeats in which the a and d position contain hydrophobic residues generate amphipathic alpha-helices which give rise to helical bundles or coiled-coil structures in proteins. Thus, SPAM is the first example of a P. falciparum antigen in which a repetitive sequence has features characteristic of a well-defined structural element. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Vic., Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7891748 ID - 191 ER - TY - JOUR AU - Marshall, V. M. AU - Anthony, R. L. AU - Bangs, M. J. AU - Purnomo AU - Anders, R. F. AU - Coppel, R. L. PY - 1994 TI - Allelic variants of the Plasmodium falciparum merozoite surface antigen 2 (MSA-2) in a geographically restricted area of Irian Jaya SP - 13-21 N1 - Jan JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 63 IS - 1 SN - 0166-6851 (Print) N1 - Allelic variants of the Plasmodium falciparum merozoite surface antigen 2 (MSA-2) in a geographically restricted area of Irian Jaya N1 - 8183312 N1 - Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Netherlands N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence DNA, Protozoan/genetics Humans Indonesia Malaria, Falciparum/parasitology Molecular Sequence Data Plasmodium falciparum/*genetics/immunology/isolation & purification Protozoan Proteins/*genetics/immunology Sequence Homology, Amino Acid *Variation (Genetics) N2 - Blood samples were collected from 12 residents of 4 villages in the Oksibil area of Irian Jaya. Eleven patients were positive for Plasmodium falciparum infection as evidenced by successful amplification of the MSA-2 gene by the polymerase chain reaction. Two patients showed evidence of infection by 2 strains of Plasmodium falciparum. All MSA-2 genes were completely sequenced and all could be assigned to one of the two major allelic families of MSA-2, however all MSA-2 gene sequences differed from previously described alleles. Five new allelic forms were identified, one of which was present in 8 of the 11 patients. Within small natural populations of P. falciparum, it appears that variation in MSA-2 approximates that seen world-wide. All samples were also analysed by hybridisation of amplified DNA to family specific probes and all samples hybridised to known probes. Our results demonstrate that there is a degree of microheterogeneity of MSA-2 that is undetectable by hybridisation studies alone. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8183312 ID - 197 ER - TY - JOUR AU - Collins, W. E. AU - Pye, D. AU - Crewther, P. E. AU - Vandenberg, K. L. AU - Galland, G. G. AU - Sulzer, A. J. AU - Kemp, D. J. AU - Edwards, S. J. AU - Coppel, R. L. AU - Sullivan, J. S. AU - et al. PY - 1994 TI - Protective immunity induced in squirrel monkeys with recombinant apical membrane antigen-1 of Plasmodium fragile SP - 711-9 N1 - Dec JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 51 IS - 6 SN - 0002-9637 (Print) N1 - Protective immunity induced in squirrel monkeys with recombinant apical membrane antigen-1 of Plasmodium fragile N1 - 7810803 N1 - Journal Article Research Support, U.S. Gov't, Non-P.H.S. United states N1 - eng KW - Adjuvants, Immunologic Animals Antibodies, Protozoan/blood Antigens, Protozoan/genetics/*immunology Antigens, Surface/genetics/immunology DNA, Protozoan/blood Disease Models, Animal Fluorescent Antibody Technique Immunization Immunization, Secondary Malaria/*prevention & control *Malaria Vaccines/adverse effects/genetics Malaria, Falciparum/prevention & control Membrane Proteins/genetics/*immunology Parasitemia/prevention & control Plasmodium/genetics/*immunology Polymerase Chain Reaction Protozoan Proteins/genetics/*immunology Recombinant Proteins/genetics/immunology Saimiri Vaccines, Synthetic/adverse effects/genetics N2 - Saimiri sciureus boliviensis monkeys were immunized with the Plasmodium fragile form of the merozoite apical membrane antigen-1 produced using the baculovirus expression system and combined with Montanide ISA 720 adjuvant. Following three immunizations, monkeys were challenged with 10,000 P. fragile trophozoite parasites. Antibody titers determined by fluorescence microscopy indicated an enhanced response following the second immunization. Four of five control animals had parasite counts > 5% 18-26 days following challenge. Four of five immunized monkeys had reduced levels of maximum parasitemia or delays in accumulated parasite counts, suggestive of protection. Rechallenge of the animals with P. falciparum resulted in three of four adjuvant control animals developing patent parasitemia whereas none of five immunized animals were infected, suggesting some level of heterologous protection. AD - Division of Parasitic Diseases, Centers for Disease Prevention & Control, Atlanta, Georgia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7810803 ID - 189 ER - TY - JOUR AU - Chaiyaroj, S. C. AU - Thompson, J. K. AU - Coppel, R. L. AU - Brown, G. V. PY - 1994 TI - Gametocytogenesis occurs in Plasmodium falciparum isolates carrying a chromosome 9 deletion SP - 163-5 N1 - Jan JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 63 IS - 1 SN - 0166-6851 (Print) N1 - Gametocytogenesis occurs in Plasmodium falciparum isolates carrying a chromosome 9 deletion N1 - 8183317 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Chromosome Deletion Chromosome Mapping Genes, Protozoan Molecular Probes Plasmodium falciparum/*genetics/*growth & development AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8183317 ID - 196 ER - TY - JOUR AU - Chaiyaroj, S. C. AU - Coppel, R. L. AU - Novakovic, S. AU - Brown, G. V. PY - 1994 TI - Multiple ligands for cytoadherence can be present simultaneously on the surface of Plasmodium falciparum-infected erythrocytes SP - 10805-8 N1 - Nov 8 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 91 IS - 23 SN - 0027-8424 (Print) N1 - Multiple ligands for cytoadherence can be present simultaneously on the surface of Plasmodium falciparum-infected erythrocytes N1 - 7526380 N1 - Dk 32094-10/dk/niddk In Vitro Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Animals Antigens, CD/metabolism Antigens, CD36 *Cell Adhesion Cell Adhesion Molecules/chemistry Cells, Cultured Erythrocyte Membrane/*metabolism Erythrocytes/*parasitology Hexosaminidases/pharmacology Humans Intercellular Adhesion Molecule-1/metabolism Ligands Molecular Weight Plasmodium falciparum Trypsin/pharmacology N2 - A major virulence factor of Plasmodium falciparum is the adherence of parasitized erythrocytes to the wall of postcapillary venules via a specific interaction between parasite-derived erythrocyte surface ligands and receptors on endothelial cells. To study this phenomenon in vitro, we selected a parasite population that expressed at least two different ligands and demonstrated that parasitized cells may coexpress ligands with specificity for multiple receptors. This selected parasite line had several antigenic and cytoadherence characteristics that were different from those of the parent line. Single parasitized erythrocytes were able to adhere to three distinct receptors via at least two separate ligands; a trypsin-sensitive molecule mediated cytoadherence to CD36 and intercellular adhesion molecule 1 and a trypsin-insensitive molecule(s) was responsible for adherence to a third receptor on the surface of melanoma cells. We present evidence that this newly discovered receptor for cytoadherence is an N-linked glycosaminoglycan, as treatment of melanoma cells with endoglycosidase H abolished cytoadherence. These observations emphasize the adaptability of P. falciparum and the complexity of the cytoadherence phenomenon. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7526380 ID - 190 ER - TY - JOUR AU - Chaiyaroj, S. C. AU - Coppel, R. L. AU - Magowan, C. AU - Brown, G. V. PY - 1994 TI - A Plasmodium falciparum isolate with a chromosome 9 deletion expresses a trypsin-resistant cytoadherence molecule SP - 21-30 N1 - Sep JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 67 IS - 1 SN - 0166-6851 (Print) N1 - A Plasmodium falciparum isolate with a chromosome 9 deletion expresses a trypsin-resistant cytoadherence molecule N1 - 7838180 N1 - Dk 32094-10/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Animals CHO Cells Cell Adhesion/genetics Cell Adhesion Molecules/*genetics Cell Line *Chromosome Deletion Chromosome Mapping Cricetinae Endothelium, Vascular/parasitology Erythrocytes/parasitology Genes, Protozoan Humans Malaria/etiology Phenotype Plasmodium falciparum/*genetics/isolation & purification/pathogenicity Protozoan Proteins/genetics Transfection Trypsin/pharmacology N2 - Sequestration of Plasmodium falciparum infected erythrocytes in the cerebral circulation is strongly implicated in the pathogenesis of cerebral malaria. From previous studies it was postulated that genes essential for cytoadherence were located on the right arm of chromosome 9 as P. falciparum isolates with a deletion in this region lost the capacity to cytoadhere in vitro and no longer expressed Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP-1) on the surface of the infected cells. We have selected a P. falciparum isolate from Papua New Guinea for high levels of cytoadherence to human umbilical vein endothelial cells (HUVECs) and have shown that the cloned parasite has several novel properties related to cytoadherence. The cloned parasite adheres to HUVECs, does not bind to melanoma cells, and expresses a surface molecule with most of the properties of PfEMP-1, despite a deletion in the right arm of chromosome 9. Interestingly, the surface expressed PfEMP-1 in this strain is resistant to trypsin treatment and infected cells continue to cytoadhere after trypsin digestion at a concentration of 100 micrograms ml-1. The receptor on HUVECs for the cloned parasite lines is a molecule different from any previously described, as parasitized cells do not adhere to soluble intercellular adhesion molecule 1, thrombospondin, vascular cell adhesion molecule 1, E-selectin or P-selectin, nor to CD36. Our work, taken together with the results from previous studies, suggest that the ability of parasites to cytoadhere is encoded in at least two distinct genomic locations in the parasite, and the diversity of receptor-ligand interaction is greater than previously described. AD - Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7838180 ID - 193 ER - TY - JOUR AU - Cha, S. AU - Leung, P. S. AU - Coppel, R. L. AU - Van de Water, J. AU - Ansari, A. A. AU - Gershwin, M. E. PY - 1994 TI - Heterogeneity of combinatorial human autoantibodies against PDC-E2 and biliary epithelial cells in patients with primary biliary cirrhosis SP - 574-83 N1 - Sep JF - Hepatology JO - Hepatology (Baltimore, Md VL - 20 IS - 3 SN - 0270-9139 (Print) N1 - Heterogeneity of combinatorial human autoantibodies against PDC-E2 and biliary epithelial cells in patients with primary biliary cirrhosis N1 - 7521314 N1 - Ai 31585/ai/niaid Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Autoantibodies/*immunology Bile Ducts/*immunology/pathology Cloning, Molecular Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/immunology/pathology Epitopes Female Humans Immunoglobulin Fab Fragments/analysis/chemistry Isomerism Liver Cirrhosis, Biliary/*immunology/*pathology Male Mutation Pyruvate Dehydrogenase Complex/chemistry/genetics/*immunology Recombinant Proteins N2 - The polyclonal nature of antimitochondrial autoantibodies and the limited success of generating human monoclonal antibodies have made analysis of fine specificity and antibody heterogeneity difficult to define. The major autoantigen of primary biliary cirrhosis is the E2 component of the pyruvate dehydrogenase pathway (PDC-E2). To address the relative importance of the region(s) in the PDC-E2 inner lipoyl domain to antibody binding, we report herein detailed profiles of 12 PDC-E2-specific antigen-binding fragments, SP1 through SP12, derived by screening of a combinatorial immunoglobulin library (derived from a primary biliary cirrhosis patient) with full-length native PDC-E2. All antigen-binding fragments are IgG isotypes and include a similar number of lambda- and kappa-chains. The antigen-binding fragments react specifically to PDC-E2 with high affinity (kappa a = 10(-7) to 10(-10) mol/L-1) and recognize a conformational epitope in the inner lipoyl domain of PDC-E2. Furthermore, the antibodies demonstrate substantial heterogeneity in recognition of different recombinant PDC-E2 fragments and differential recognition patterns against mutant constructs of the human PDC-E2 inner lipoyl domain (amino acid residues 91 to 227). In addition, five of the antigen-binding fragment clones (SP1, 3, 4, 8 and 12) demonstrate different staining patterns on biliary epithelial cells of patients with primary biliary cirrhosis but not control liver disease; some antigen-binding fragments specifically stained the apical region of biliary epithelium, a pattern distinct from that of typical mitochondrial staining. The response to the inner lipoyl domain is not, however, monospecific, and there is much more heterogeneity in fine specificity than could be accounted for by arbitrary reshuffling of variable immunoglobulin heavy and light chains into unnatural combinations. AD - Division of Rheumatology, Allergy and Clinical Immunology, School of Medicine, University of California at Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7521314 ID - 194 ER - TY - JOUR AU - Van de Water, J. AU - Turchany, J. AU - Leung, P. S. AU - Lake, J. AU - Munoz, S. AU - Surh, C. D. AU - Coppel, R. AU - Ansari, A. AU - Nakanuma, Y. AU - Gershwin, M. E. PY - 1993 TI - Molecular mimicry in primary biliary cirrhosis. Evidence for biliary epithelial expression of a molecule cross-reactive with pyruvate dehydrogenase complex-E2 SP - 2653-64 N1 - Jun JF - J Clin Invest JO - The Journal of clinical investigation VL - 91 IS - 6 SN - 0021-9738 (Print) N1 - Molecular mimicry in primary biliary cirrhosis. Evidence for biliary epithelial expression of a molecule cross-reactive with pyruvate dehydrogenase complex-E2 N1 - 8514873 N1 - Dk-39588/dk/niddk Dk-6-2274/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Antibodies, Monoclonal Autoantibodies/analysis Bile/chemistry Bile Ducts/enzymology/*metabolism Cross Reactions Dihydrolipoyllysine-Residue Acetyltransferase Epithelium/enzymology/metabolism Humans Immunoglobulin A/analysis Immunoglobulin G/analysis Immunoglobulin Isotypes/analysis Immunohistochemistry Liver Cirrhosis, Biliary/immunology/*metabolism Microscopy/methods Mitochondria, Heart/enzymology/immunology Pyruvate Dehydrogenase Complex/immunology/metabolism N2 - Sera from patients with primary biliary cirrhosis (PBC) react with enzymes of the 2-oxo dehydrogenase pathways, particularly PDC-E2. These enzymes are present in all nucleated cells, yet autoimmune damage is confined to biliary epithelial cells. Using a panel of eight mouse monoclonal antibodies and a human combinatorial antibody specific for PDC-E2, we examined by indirect immunofluorescence and confocal microscopy sections of liver from patients with PBC, progressive sclerosing cholangitis, and hepatocarcinoma. The monoclonal antibodies gave typical mitochondrial immunofluorescence on biliary epithelium and on hepatocytes from patients with either PBC, progressive sclerosing cholangitis, or hepatocarcinoma. However, one of eight mouse monoclonal antibodies (C355.1) and the human combinatorial antibody reacted with great intensity and specificity with the luminal region of biliary epithelial cells from patients with PBC. Simultaneous examination of these sections with an antiisotype reagent for human IgA revealed high IgA staining in the luminal region of biliary epithelial cells in patients with PBC. IgG and IgA antibodies to PDC-E2 were detected in the bile of patients with PBC but not normal controls. We believe that this data may be interpreted as indicating that a molecule cross-reactive with PDC-E2 is expressed at high levels in the luminal region of biliary epithelial cells in PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, School of Medicine, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8514873 ID - 201 ER - TY - JOUR AU - Marshall, V. M. AU - Coppel, R. L. PY - 1993 TI - Antigenic typing of field isolates of Plasmodium by DNA techniques SP - 223-48 JF - Methods Mol Biol JO - Methods in molecular biology (Clifton, N.J VL - 21 SN - 1064-3745 (Print) N1 - Antigenic typing of field isolates of Plasmodium by DNA techniques N1 - 8220718 N1 - Journal Article United states N1 - eng KW - Alleles Animals Antigens, Protozoan/*genetics Automation Blood/parasitology Chromatography/methods DNA, Protozoan/*analysis Fluorescent Dyes *Genes, Protozoan Humans Malaria/classification/*parasitology Parasitology/*methods Plasmodium/*classification/genetics/immunology/isolation & purification Polymerase Chain Reaction/*methods Protozoan Proteins/genetics/immunology Sequence Analysis, DNA/*methods AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8220718 ID - 205 ER - TY - JOUR AU - Leung, P. S. AU - Watanabe, Y. AU - Munoz, S. AU - Teuber, S. S. AU - Patel, M. S. AU - Korenberg, J. R. AU - Hara, P. AU - Coppel, R. AU - Gershwin, M. E. PY - 1993 TI - Chromosome localization and RFLP analysis of PDC-E2: the major autoantigen of primary biliary cirrhosis SP - 335-40 JF - Autoimmunity JO - Autoimmunity VL - 14 IS - 4 SN - 0891-6934 (Print) N1 - Chromosome localization and RFLP analysis of PDC-E2: the major autoantigen of primary biliary cirrhosis N1 - 8102256 N1 - 1-dk-6-2774/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Switzerland N1 - eng KW - Autoantigens/*genetics *Chromosome Mapping Chromosomes, Human, Pair 11 Genotype Humans Liver Cirrhosis, Biliary/genetics/*immunology *Polymorphism, Restriction Fragment Length Pyruvate Dehydrogenase Complex/*genetics N2 - Patients with primary biliary cirrhosis are well known for the presence of titer antibodies against dihydrolipoamide acetyltransferase, the E2 subunit of the pyruvate dehydrogenase complex. We have taken advantage of a cDNA probe for dihydrolipoamide acetyltransferase to explore the possibility of polymorphism of the E2 subunit by probing genomic DNA from 38 patients with primary biliary cirrhosis and 26 healthy controls. To detect restriction fragment length polymorphism, DNA was digested with ten specific restriction enzymes that often detect polymorphism, including Bam HI, Bgl II, Eco RI, Hind III, Hinf I, Msp I, Pst I, Pvu II, Rsa I and Taq I. A Taq I polymorphism was found in 19 of 38 patients with PBC and 6 of 26 normal controls. In addition, using fluorescence in situ hybridization, the gene for dihydrolipoamide acetyltransferase was mapped on human chromosome 11 band q23.1. Interestingly, this region of the long arm of chromosome 11 is often associated with cytogenetic abnormalities, including translocations. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8102256 ID - 206 ER - TY - JOUR AU - Leedman, P. J. AU - Faulkner-Jones, B. AU - Cram, D. S. AU - Harrison, P. J. AU - West, J. AU - O'Brien, E. AU - Simpson, R. AU - Coppel, R. L. AU - Harrison, L. C. PY - 1993 TI - Cloning from the thyroid of a protein related to actin binding protein that is recognized by Graves disease immunoglobulins SP - 5994-8 N1 - Jul 1 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 90 IS - 13 SN - 0027-8424 (Print) N1 - Cloning from the thyroid of a protein related to actin binding protein that is recognized by Graves disease immunoglobulins N1 - 8327473 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Base Sequence *Cloning, Molecular Graves Disease/*immunology Humans Immunoglobulins/*immunology Mice Microfilament Proteins/chemistry/*genetics/immunology/physiology Molecular Sequence Data RNA, Messenger/analysis Receptors, Thyrotropin/chemistry/physiology Recombinant Fusion Proteins/chemistry Sequence Homology, Amino Acid Signal Transduction Thyroid Gland/*chemistry N2 - Human actin binding protein (ABP) links specific membrane glycoproteins to cytoskeletal actin microfilaments. In human platelets and leukocytes, ABP directly links, respectively, the membrane glycoproteins GPIb and the high-affinity Fc receptor for IgG (Fc gamma IR) to cytoskeletal actin microfilaments. Similar interaction between the thyrotropin (TSH) receptor and ABP in endocrine cells might explain the rapid and profound disruption of actin microfilaments induced by TSH in cultured thyroid follicular cells. By screening a thyroid lambda gt11 cDNA expression library with serum from a Graves disease patient, we identified a clone encoding a protein, designated truncated ABP (TABP), that shares extensive homology (approximately 70%) with ABP. TABP is a truncated ABP-like protein with an open reading frame of 195 aa that encodes a protein of approximately 21 kDa. TABP lacks an actin binding domain but contains two predicted beta-sheet repeats within which is a putative dimerization domain and between which lies a putative glycoprotein binding site containing a consensus site for phosphorylation by Ca(2+)-calmodulin kinase II. TABP contains a unique C-terminal insertion within which lies a hydrophobic predicted membrane-associated region, absent from ABP. Although TABP mRNA is expressed widely, immunoblot analysis demonstrated the presence of TABP antibodies specifically in the sera of a minority of subjects with autoimmune thyroid disease. A 24-residue sequence of similarity was identified between the TSH receptor and platelet glycoprotein GPIb alpha that may represent a transmembrane ABP binding site. We suggest, therefore, that signal transduction by TSH in the thyroid involves direct linkage of the TSH receptor to actin microfilaments by ABP and that TABP may interact with ABP to mediate TSH-induced actin microfilament disruption. AD - Burnet Clinical Research Unit, Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8327473 ID - 200 ER - TY - JOUR AU - Cram, D. S. AU - Fisicaro, N. AU - McNeilage, L. J. AU - Coppel, R. L. AU - Harrison, L. C. PY - 1993 TI - Antibody specificities of Thai and Australian scleroderma sera with topoisomerase I recombinant fusion proteins SP - 6872-81 N1 - Dec 15 JF - J Immunol VL - 151 IS - 12 SN - 0022-1767 (Print) N1 - Antibody specificities of Thai and Australian scleroderma sera with topoisomerase I recombinant fusion proteins N1 - 7505017 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't United states 1950) N1 - eng KW - Amino Acid Sequence Antibody Specificity Australia Autoantibodies/*blood Autoantigens/genetics Base Sequence Cloning, Molecular DNA Primers/genetics DNA Topoisomerases, Type I/genetics/*immunology DNA, Complementary/genetics Epitopes/genetics Humans Molecular Sequence Data Peptide Mapping Recombinant Fusion Proteins/genetics/immunology Scleroderma, Systemic/*enzymology/*immunology Thailand N2 - Autoantibodies that react with the nuclear enzyme topoisomerase I (Topo I) are used as a diagnostic marker of diffuse scleroderma. To better define immune reactivity to Topo I, antibody epitopes in two patient populations were analyzed using recombinant Topo I proteins. Two overlapping partial cDNA clones encoding the complete amino acid sequence of Topo I were isolated from human placenta. Using the polymerase chain reaction, specific regions of Topo I were amplified and cloned into the pGEX expression vectors. To map Topo I epitopes, recombinant fusion proteins were analyzed by immunoblotting with 66 anti-Topo I sera from Thai and Australian patients with diffuse scleroderma. Six distinct epitope regions were identified along the length of the 765 amino acid enzyme. Almost all sera contained antibodies that recognized the midregion of Topo I (amino acids 453-560), as well as antibodies to one of more of the other epitope regions. Sixty percent of the sera contained antibodies that recognized a COOH-terminal epitope region (amino acids 658-765) encompassing the active site of the enzyme. This subset of Topo I antibodies could be responsible for the inhibition of enzymatic activity previously reported in vitro. Heterogeneous patterns of reactivity with the six Topo I epitope regions were observed, although over half the sera could be assigned to one of six distinct patterns. In general, antibodies in the Thai sera reacted more strongly with the six epitope regions. Furthermore, two of the epitope regions reacted exclusively with Thai sera, suggesting a degree of racial or geographical specificity in the autoantibody response to Topo I. The identification of multiple epitopes in Topo I conforms with the polyclonal autoantibody response to intracellular Ag found in other multisystem autoimmune diseases and is presumed to be driven by the presentation of multiple peptides from Topo I itself. AD - Burnet Clinical Research Unit, Walter and Eliza Hall Institute of Medical Research, Parkville, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7505017 ID - 198 ER - TY - JOUR AU - Coppel, R. L. AU - Smith, F. M. AU - Petersen, C. PY - 1993 TI - Antibody screening of expression libraries SP - 277-96 JF - Methods Mol Biol JO - Methods in molecular biology (Clifton, N.J VL - 21 SN - 1064-3745 (Print) N1 - Antibody screening of expression libraries N1 - 8220721 N1 - Journal Article United states N1 - eng KW - Animals Antibodies/*immunology Antibody Specificity Antigens/*genetics/immunology Antigens, Helminth/genetics/immunology Antigens, Protozoan/genetics/immunology Cloning, Molecular DNA, Complementary/genetics *Gene Library Genes Genes, Protozoan Genetic Vectors Immunoassay Parasites/genetics/*immunology Parasitology/*methods Recombinant Fusion Proteins/genetics/*immunology AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8220721 ID - 204 ER - TY - JOUR AU - Cha, S. AU - Leung, P. S. AU - Gershwin, M. E. AU - Fletcher, M. P. AU - Ansari, A. A. AU - Coppel, R. L. PY - 1993 TI - Combinatorial autoantibodies to dihydrolipoamide acetyltransferase, the major autoantigen of primary biliary cirrhosis SP - 2527-31 N1 - Mar 15 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 90 IS - 6 SN - 0027-8424 (Print) N1 - Combinatorial autoantibodies to dihydrolipoamide acetyltransferase, the major autoantigen of primary biliary cirrhosis N1 - 8460168 N1 - Dk39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acetyltransferases/*immunology Antigen-Antibody Complex Autoantibodies/*genetics Autoantigens/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Enzyme-Linked Immunosorbent Assay Female Fluorescent Antibody Technique Gene Library Humans Immunoglobulin Fab Fragments/genetics Immunoglobulin G/genetics Kinetics Liver/pathology Liver Cirrhosis, Biliary/enzymology/*immunology/surgery Liver Transplantation Lymph Nodes/*immunology *Pyruvate Dehydrogenase Complex RNA, Messenger/genetics/isolation & purification Tumor Cells, Cultured N2 - mRNA from a regional lymph node of a patient with primary biliary cirrhosis (PBC) was used to construct a combinatorial immunoglobulin library in the lambda phage vector system. Six human monoclonal IgG Fab clones (LC1-LC6) specific for the major autoantigen of PBC--dihydrolipoamide acetyltransferase, the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2)--were isolated, appearing at a frequency of 0.01% in the combinatorial immunoglobulin library. These Fab clones recognize human PDC-E2 with high affinity (Ka = 10(-7)-10(-9) M-1). Using both immunoblotting and ELISA, LC1-LC6 showed little cross-reactivity to any of the other autoantigens commonly recognized by PBC sera or to other antigens commonly recognized by PBC sera or to other antigens such as histone, calf thymus DNA, and bovine serum albumin. The Fab monoclonal antibodies show a typical anti-mitochondrial staining pattern in HEp-2 cells but react strongly with the luminal aspect of biliary epithelial cells of patients with PBC. Our results demonstrate that a recombinant combinatorial immunoglobulin library can be used to isolate high-affinity Fabs against a specific autoantigen. Such reagents will facilitate the analysis of immunoglobulin gene structure, idiotype, and antigen-antibody interactions in autoimmune disease. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8460168 ID - 203 ER - TY - JOUR AU - Bickle, Q. AU - Anders, R. F. AU - Day, K. AU - Coppel, R. L. PY - 1993 TI - The S-antigen of Plasmodium falciparum: repertoire and origin of diversity SP - 189-96 N1 - Oct JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 61 IS - 2 SN - 0166-6851 (Print) N1 - The S-antigen of Plasmodium falciparum: repertoire and origin of diversity N1 - 8264723 N1 - Comparative Study Journal Article Netherlands N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/analysis/*biosynthesis/genetics Antigens, Surface/analysis/*biosynthesis/genetics Base Sequence Consensus Sequence DNA Primers Molecular Sequence Data Plasmodium falciparum/genetics/*immunology Polymerase Chain Reaction Polymorphism, Genetic Repetitive Sequences, Nucleic Acid Sequence Homology, Amino Acid *Variation (Genetics) N2 - S-antigens are heat-stable, highly polymorphic proteins released by Plasmodium falciparum at the time of schizont rupture. Previously determined S-antigen sequences allowed the proposal of a general gene structure consisting of 5 sequence blocks. The sequence of the central block of tandem repeats provides a useful means of distinguishing the S-antigen allele and also its serotype, whereas the amino and carboxy terminal sequences defined the S-antigen family, 4 of which have been described. We present the sequence of 3 new S-antigen alleles, for the isolates HB3, KF1916 and KF1917. The allele-defining repeat sequence is ETGPGKAGEQG for HB3, GDQTEGS(S/A)GGK for KF1917 and AGSNE(E/K) for KF1916. The sequences of these newly described S-antigens are consistent with the proposed general gene structure and all belong to defined families, although carboxy-terminal sequences appear to be much more variable within a family than previously realised. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8264723 ID - 199 ER - TY - JOUR AU - Anders, R. F. AU - McColl, D. J. AU - Coppel, R. L. PY - 1993 TI - Molecular variation in Plasmodium falciparum: polymorphic antigens of asexual erythrocytic stages SP - 239-53 N1 - May JF - Acta Trop JO - Acta tropica VL - 53 IS - 3-4 SN - 0001-706X (Print) N1 - Molecular variation in Plasmodium falciparum: polymorphic antigens of asexual erythrocytic stages N1 - 8100673 N1 - Journal Article Review Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigenic Variation/*genetics Antigens, Protozoan/*genetics Erythrocytes/parasitology Genes, Protozoan/*genetics Humans Molecular Sequence Data Plasmodium falciparum/*genetics/growth & development/immunology Polymorphism, Genetic/genetics N2 - Numerous polymorphic antigens of the asexual erythrocytic stages of P. falciparum are now well characterized. Diversity in some of these antigens, including MSA-1, MSA-2 and the S-antigen is associated with changes in the repeat sequences which are frequently a prominent structural feature of malaria antigens. It is not known whether the variation in repeats causes allelic gene products to adopt different conformations but variation in and around the repeats in SPAM, a newly characterized secreted antigen, preserve the unusual alanine-heptad repeats which we assume generate a helical bundle in this protein. Mutations in non-repetitive regions of the S-antigen and in AMA-1, an antigen lacking repeats, are strongly biased towards those which alter the amino acid sequence. This and other evidence indicates the operation of biological selection but the role of immune responses as a selection pressure operating on these diverse antigens remains to be established. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8100673 ID - 202 ER - TY - JOUR AU - Marshall, V. M. AU - Coppel, R. L. AU - Anders, R. F. AU - Kemp, D. J. PY - 1992 TI - Two novel alleles within subfamilies of the merozoite surface antigen 2 (MSA-2) of Plasmodium falciparum SP - 181-4 N1 - Jan JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 50 IS - 1 SN - 0166-6851 (Print) N1 - Two novel alleles within subfamilies of the merozoite surface antigen 2 (MSA-2) of Plasmodium falciparum N1 - 1542312 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - *Alleles Amino Acid Sequence Animals *Antigens, Protozoan Antigens, Surface/*genetics Base Sequence DNA, Protozoan Molecular Sequence Data Plasmodium falciparum/*genetics Protozoan Proteins/*genetics Sequence Alignment AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1542312 ID - 213 ER - TY - JOUR AU - Leung, P. S. AU - Iwayama, T. AU - Prindiville, T. AU - Chuang, D. T. AU - Ansari, A. A. AU - Wynn, R. M. AU - Dickson, R. AU - Coppel, R. AU - Gershwin, M. E. PY - 1992 TI - Use of designer recombinant mitochondrial antigens in the diagnosis of primary biliary cirrhosis SP - 367-72 N1 - Mar JF - Hepatology JO - Hepatology (Baltimore, Md VL - 15 IS - 3 SN - 0270-9139 (Print) N1 - Use of designer recombinant mitochondrial antigens in the diagnosis of primary biliary cirrhosis N1 - 1371979 N1 - 1-dk-6-2274/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Autoantibodies/analysis Autoantigens/chemistry/*diagnostic use Drug Design Enzyme-Linked Immunosorbent Assay Epitopes Humans *Immunologic Tests Ketone Oxidoreductases/chemistry Liver Cirrhosis, Biliary/*diagnosis Mitochondria/*immunology Multienzyme Complexes/chemistry Pyruvate Dehydrogenase Complex/chemistry Recombinant Proteins Sensitivity and Specificity N2 - The appearance of autoantibodies against mitochondria in patients with primary biliary cirrhosis has been known for more than 25 yr. In the past, based on the biochemical complexity of the mitochondrion and the use of crude extracts for immunodiagnosis, a degree of nonspecificity in assaying for antibodies to mitochondria has been present. This problem has been largely circumvented by the cloning of the mitochondrial antigens and the identification of the E2 subunits of the pyruvate dehydrogenase complex and the branched chain 2-oxo-acid dehydrogenase complex as the major and immunodominant autoantigens of primary biliary cirrhosis. More than 90% of patients with primary biliary cirrhosis have been shown to react with one or both of these enzymes using either recombinant antigen or purified native protein. Approximately 10% of patients recognize only E2 subunits of branched chain 2-oxo-acid dehydrogenase complex and not pyruvate dehydrogenase complex. Such patients would be missed by diagnostic assay that has a low sensitivity to antibodies against E2 subunits of branched chain 2-oxo-acid dehydrogenase complex. The use of recombinant and biochemically pure antigens has permitted structural and conformational analysis of epitope mapping. We have taken advantage of the antigenic mapping studies of both primary biliary cirrhosis and branched chain 2-oxo-acid dehydrogenase complex E2 subunits and designed a molecule that expresses the immunodominant epitopes of both. Using this dual-headed molecule that coexpresses the epitope of two different antigens, we report herein a sensitive and reproducible assay for antibodies to mitochondria in patients with primary biliary cirrhosis.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Department of Internal Medicine, University of California, Davis. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1371979 ID - 211 ER - TY - JOUR AU - Iwayama, T. AU - Leung, P. S. AU - Rowley, M. AU - Munoz, S. AU - Nishioka, M. AU - Nakagawa, T. AU - Dickson, E. R. AU - Coppel, R. L. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 1992 TI - Comparative immunoreactive profiles of Japanese and American patients with primary biliary cirrhosis against mitochondrial autoantigens SP - 28-33 JF - Int Arch Allergy Immunol JO - International archives of allergy and immunology VL - 99 IS - 1 SN - 1018-2438 (Print) N1 - Comparative immunoreactive profiles of Japanese and American patients with primary biliary cirrhosis against mitochondrial autoantigens N1 - 1483064 N1 - Dk 39588/dk/niddk Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. Switzerland N1 - eng KW - Antigen-Antibody Reactions/*immunology Autoantibodies/immunology Autoantigens/*immunology Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Humans Immunoblotting Incidence Japan/ethnology Liver Cirrhosis, Biliary/ethnology/*immunology Mitochondria/*immunology Mitochondria, Heart/immunology Pyruvate Dehydrogenase Complex/immunology Random Allocation United States/ethnology N2 - Primary biliary cirrhosis (PBC) has been described among various ethnic and racial populations in all parts of the world. However, the incidence and prevalence of PBC varies considerably in different geographic areas. It has the highest frequency in Northern Europe, is considerably lower in Japan and still lower in other parts of Asia. There has not hitherto been a detailed immunological profile of antimitochondrial antibodies according to geographic region. We have used recombinant or purified preparations from the 2-oxo-acid dehydrogenase enzyme complexes, the major mitochondrial autoantigens in PBC (PDC-E2, BCOADC-E2, OGDC, protein X and PDC-E1 alpha) to compare the reactivity of sera from either similarly staged sera from Japanese (n = 23) or American-Caucasian patients (n = 39) with PBC. In all cases, the first available sera following diagnosis was selected. Interestingly, only 65% of Japanese patients reacted by ELISA with PDC-E2 compared with more than 95% of the North American group. Moreover, the level of enzyme-inhibitory antibodies to PDC was lower in the Japanese. Our findings prompt the need for characterization of specific susceptibility genes and environmental factors in various parts of the world to clarify the etiology of PBC. AD - First Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Japan. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1483064 ID - 214 ER - TY - JOUR AU - Gershwin, M. E. AU - Rowley, M. AU - Davis, P. A. AU - Leung, P. AU - Coppel, R. AU - Mackay, I. R. PY - 1992 TI - Molecular biology of the 2-oxo-acid dehydrogenase complexes and anti-mitochondrial antibodies SP - 47-61 JF - Prog Liver Dis JO - Progress in liver diseases VL - 10 SN - 1060-913X (Print) N1 - Molecular biology of the 2-oxo-acid dehydrogenase complexes and anti-mitochondrial antibodies N1 - 1296237 N1 - Journal Article Review United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) *Autoantibodies Autoantigens/chemistry Humans Ketone Oxidoreductases/antagonists & inhibitors/chemistry/*immunology Liver Cirrhosis, Biliary/enzymology/genetics/immunology Mitochondria/enzymology/immunology Molecular Structure Multienzyme Complexes/antagonists & inhibitors/chemistry/*immunology T-Lymphocytes/immunology N2 - The confluence of molecular biology and clinical medicine has provided new and valuable insights in PBC. Our understanding of the immunobiology of PBC has changed dramatically using this new technology. It is now possible to explicitly define mitochondrial autoantigens and examine recognition sites, by using autoantibodies, at the primary sequence level. In addition, cloned antigens have been developed to reliably assay for presence of autoantibodies; the use of cloned recombinant antigens should replace that of traditional immunofluorescence for AMA assay. It is also now possible to begin the task of defining the role of T cells in the immunopathology of PBC and exploring the issue of whether immunotherapy is possible. Finally, there is increasing evidence that PDC-E2 is located on the cell membrane of biliary epithelial cells. The mechanism for this expression remains to be studied. The explosion of data in PBC is an example of the serendipity and synergy brought about by application of new techniques to investigate old problems. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California, Davis. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1296237 ID - 215 ER - TY - JOUR AU - Davis, P. A. AU - Leung, P. AU - Manns, M. AU - Kaplan, M. AU - Munoz, S. J. AU - Gorin, F. A. AU - Dickson, E. R. AU - Krawitt, E. AU - Coppel, R. AU - Gershwin, M. E. PY - 1992 TI - M4 and M9 antibodies in the overlap syndrome of primary biliary cirrhosis and chronic active hepatitis: epitopes or epiphenomena? SP - 1128-36 N1 - Nov JF - Hepatology JO - Hepatology (Baltimore, Md VL - 16 IS - 5 SN - 0270-9139 (Print) N1 - M4 and M9 antibodies in the overlap syndrome of primary biliary cirrhosis and chronic active hepatitis: epitopes or epiphenomena? N1 - 1330864 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Adenoma, Bile Duct/immunology Autoantibodies/*blood Autoantigens/immunology Bile Duct Neoplasms/immunology Cholangitis, Sclerosing/immunology Complement Fixation Tests Enzyme-Linked Immunosorbent Assay False Positive Reactions Hepatitis, Chronic/*complications/*immunology Humans Immunoblotting Liver Cirrhosis, Alcoholic/immunology Liver Cirrhosis, Biliary/*complications/*immunology Lupus Erythematosus, Systemic/immunology Oxidoreductases Acting on Sulfur Group Donors/blood/immunology Phosphorylases/blood/immunology Prognosis Scleroderma, Systemic/immunology Syndrome N2 - Before the identification of the major mitochondrial antigens of primary biliary cirrhosis as components of the 2-oxo-acid dehydrogenase enzyme family, mitochondrial autoantigens were believed to be extremely heterogeneous and were divided into nine subtypes termed M1 to M9. This classification was based on the data derived from the relatively nonspecific biochemical and immunological techniques that were available. After the cloning and definition of the major autoantigens, more than 95% of the sera of patients with primary biliary cirrhosis were found to react with components of the 2-oxo-dehydrogenase enzymes; these enzymes correspond to the old M2 classification. Two other "M" species, dubbed M4 and M9, have attracted significant attention because they have been postulated to be prognostic indicators and more recently have been tentatively identified respectively as sulfite oxidase (EC 1.8.3.1) and glycogen phosphorylase (EC 2.4.1.1). Indeed, patients with the "overlap syndrome" are reported to have antibodies to M4 and a poor prognosis, whereas patients with antibodies to M9 have a favorable prognosis. To address the significance and definition of M4 and M9, we performed in-depth studies of sera from 11 patients with the overlap syndrome, 75 patients with primary biliary cirrhosis, 19 chronic active hepatitis patients, 13 patients with primary sclerosing cholangitis, 10 patients with cholangiocarcinoma, 20 patients with systemic lupus erythematosus, 20 patients with alcoholic cirrhosis, 17 patients with scleroderma and 30 normal individuals, using techniques of ELISA, complement fixation, immunoblotting and enzyme inhibition. We report herein that we were unable to show any disease-specific reactivity toward the proposed M4 and M9 antigens.(ABSTRACT TRUNCATED AT 250 WORDS) AD - University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1330864 ID - 209 ER - TY - JOUR AU - Coppel, R. L. PY - 1992 TI - Repeat structures in a Plasmodium falciparum protein (MESA) that binds human erythrocyte protein 4.1 SP - 335-47 N1 - Feb JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 50 IS - 2 SN - 0166-6851 (Print) N1 - Repeat structures in a Plasmodium falciparum protein (MESA) that binds human erythrocyte protein 4.1 N1 - 1741020 N1 - Dk32094/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Netherlands N1 - eng KW - Amino Acid Sequence Animals Base Sequence Cloning, Molecular *Cytoskeletal Proteins DNA, Protozoan/genetics Membrane Proteins/*metabolism Molecular Sequence Data *Neuropeptides Plasmodium falciparum/*genetics Protozoan Proteins/*genetics/metabolism *Repetitive Sequences, Nucleic Acid Sequence Homology, Nucleic Acid N2 - The mature-parasite-infected erythrocyte surface antigen (MESA, also known as PfEMP-2 and pp300) of Plasmodium falciparum is a phosphoprotein of approx. 250-300 kDa that is exported from the parasite to the erythrocyte membrane skeleton where it binds to protein 4.1. Determination of the primary sequence of MESA reveals that it is encoded by 2 exons, a structure common to other exported proteins of P. falciparum. The MESA protein is heavily charged and contains 7 distinct repeat regions that compose over 60% of the protein. The predicted secondary structure suggests that MESA is a fibrillar protein and it shows similarity to a number of cytoskeletal and neurofilament proteins, including myosin, a protein that itself binds to protein 4.1. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1741020 ID - 212 ER - TY - JOUR AU - Coppel, R. L. PY - 1992 TI - Malaria--revealing the ties that bind SP - 393-4; discussion 411 N1 - Dec JF - Parasitol Today JO - Parasitology today (Personal ed VL - 8 IS - 12 SN - 0169-4758 (Print) N1 - Malaria--revealing the ties that bind N1 - 15463550 N1 - Journal Article England N1 - eng AD - Ross Coppel is at the Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3050, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15463550 ID - 207 ER - TY - JOUR AU - Caldwell, S. H. AU - Leung, P. S. AU - Spivey, J. R. AU - Prindiville, T. AU - de Medina, M. AU - Saicheur, T. AU - Rowley, M. AU - Reddy, K. R. AU - Coppel, R. AU - Jeffers, L. J. AU - et al. PY - 1992 TI - Antimitochondrial antibodies in kindreds of patients with primary biliary cirrhosis: antimitochondrial antibodies are unique to clinical disease and are absent in asymptomatic family members SP - 899-905 N1 - Oct JF - Hepatology JO - Hepatology (Baltimore, Md VL - 16 IS - 4 SN - 0270-9139 (Print) N1 - Antimitochondrial antibodies in kindreds of patients with primary biliary cirrhosis: antimitochondrial antibodies are unique to clinical disease and are absent in asymptomatic family members N1 - 1398496 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Aged Antibodies/*analysis Biopsy Enzyme-Linked Immunosorbent Assay Female Fluorescent Antibody Technique Humans Immunoblotting Liver/pathology Liver Cirrhosis, Biliary/enzymology/genetics/*immunology Middle Aged Mitochondria/*immunology Pyruvate Dehydrogenase Complex/analysis/immunology N2 - The 2-oxo-acid dehydrogenase family of enzymes have been identified as the major mitochondrial autoantigens of primary biliary cirrhosis. Using immunoblotting, enzyme-linked immunosorbent assay and enzyme inhibition with both purified mitochondrial proteins and recombinant autoantigens, we have studied family members and spouses of patients with primary biliary cirrhosis for the presence of antimitochondrial antibodies. Antimitochondrial antibodies and other common autoantigens were also tested for by indirect immunofluorescence. This study included 27 index patients with primary biliary cirrhosis, 15 spouses and 48 first- and second-degree relatives. Overall, 7 relatives (11%) were positive for autoantibodies to nuclear and cytoplasmic antigens by indirect immunofluorescence against mouse liver and stomach sections. However, with immunofluorescence, the reactivity strictly paralleled that of antimitochondrial antibodies in only one of these (1:640)--a sibling with mild pruritus and a liver biopsy specimen diagnostic of primary biliary cirrhosis despite normal levels of serum alkaline phosphatase. In addition, one of the mothers, who had a history of sarcoidosis, was positive by immunoblotting for antibodies to the E2 subunit of the pyruvate dehydrogenase complex and protein X. All other relatives were negative for all of the assays. Antibodies to neither the 2-oxo-acid dehydrogenase enzymes nor the recently proposed family of naturally occurring mitochondrial antibodies were found in spouses or healthy relatives. Three other first-degree relatives suffered from liver disease: two died (one from primary biliary cirrhosis and the other from an unknown type of liver disease) and one (a sibling with primary biliary cirrhosis) was unavailable for testing. Our results are consistent with a familial predisposition to primary biliary cirrhosis.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Center for Liver Disease, University of Miami, Florida. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1398496 ID - 210 ER - TY - JOUR AU - Bickle, Q. AU - Coppel, R. L. PY - 1992 TI - A fourth family of the Plasmodium falciparum S-antigen SP - 141-50 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 56 IS - 1 SN - 0166-6851 (Print) N1 - A fourth family of the Plasmodium falciparum S-antigen N1 - 1474992 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence Cloning, Molecular DNA, Protozoan/genetics Genes, Protozoan Molecular Sequence Data Multigene Family Plasmodium falciparum/*genetics/*immunology Polymerase Chain Reaction Protozoan Proteins/genetics Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid N2 - The S-antigen of Plasmodium falciparum is a highly diverse heat stable protein that is located in the parasitophorous vacuole of the mature asexual intraerythrocytic parasite. The gene for S-antigen exists within the parasite population as multiple alleles at a single locus. Its sequence contains a large central block of tandemly arranged peptides that are identical or very similar in one allele but differ widely in sequence, repeat length and number among different alleles, and consequently antigenic specificity. Thus, antibodies directed against the repeat region can be used to define the serotype of an S-antigen. Flanking this repeat block are 2 short regions of non-repetitive sequence which have been described as occurring in three different forms, each of which is used to define a single S-antigen family. We present the S-antigen sequence for the isolate 3D7 which defines not only a novel serotype but also a novel S-antigen family. The central repeat block is composed of 57 copies of an 8-residue peptide with consensus sequence ED(E/K)VSNG(R/G). Comparison of the four S-antigen families reveals that they differ considerably from each other with variation being most pronounced in the carboxy terminal-flanking region. This pattern of sequence variation differs considerably from that found for MSA-1 and MSA-2, the only other diverse proteins of P. falciparum for which sequence information is available. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1474992 ID - 208 ER - TY - JOUR AU - Van de Water, J. AU - Ansari, A. A. AU - Surh, C. D. AU - Coppel, R. AU - Roche, T. AU - Bonkovsky, H. AU - Kaplan, M. AU - Gershwin, M. E. PY - 1991 TI - Evidence for the targeting by 2-oxo-dehydrogenase enzymes in the T cell response of primary biliary cirrhosis SP - 89-94 N1 - Jan 1 JF - J Immunol VL - 146 IS - 1 SN - 0022-1767 (Print) N1 - Evidence for the targeting by 2-oxo-dehydrogenase enzymes in the T cell response of primary biliary cirrhosis N1 - 1984455 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Antigens, CD/analysis Antigens, Differentiation, T-Lymphocyte/analysis Bile Ducts/enzymology/immunology Fluorescent Antibody Technique Ketone Oxidoreductases/*immunology Liver Cirrhosis, Biliary/enzymology/*immunology Multienzyme Complexes/*immunology Pyruvate Dehydrogenase Complex/*immunology T-Lymphocyte Subsets/immunology T-Lymphocytes/*immunology N2 - Primary biliary cirrhosis (PBC) is a chronic autoimmune liver disease that includes the presence of lymphoid infiltrates in portal tracts, high titer autoantibodies against pyruvate dehydrogenase-E2 (PDH-E2) and branched chain ketoacid dehydrogenase-E2 (BCKD-E2), and biliary tract destruction. The mechanism by which the autoimmune response is induced, the specificity of damage to the biliary epithelium, and the role of T cells in PBC are still unknown. To address these issues, we have taken advantage of a mouse mAb, coined C355.1, and studied its reactivity against a panel of liver tissue from normal subjects as well as a panel of liver specimens from patients with PBC, progressive sclerosing cholangitis, and chronic active hepatitis (CAH). C355.1, much like human autoantibodies to PDH-E2, reacts exclusively by immunoblotting with PDH-E2, binds to the inner lipoyl domain of the protein, and inhibits PDH-E2 activity in vitro. In addition, we have also attempted to develop cloned T cell lines that react with PDH-E2 and/or BCKD-E2 using liver biopsies from patients with PBC, compared with CAH. Although monoclonal C355.1 produced typical mitochondrial fluorescence on sections of normal liver, pancreas, lung, heart, thyroid, and kidney, it produced a distinct and intense reactivity when used to stain the bile ducts of patients with PBC. Nine of 13 PBC liver biopsies studied herein contained bile ducts on light microscopy, all of which reacted intensely at a 1:100 culture supernatant dilution of monoclonal C355.1. In contrast, although bile ducts of liver specimens from normals, CAH, and progressive sclerosing cholangitis also reacted with C355.1, such reactivity was exclusively mitochondrial and readily detectable only at a dilution of 1:2. More importantly, we generated CD4+, CD8-, alpha beta TCR+ cloned T cell lines from patients with PBC, but not from CAH, that produced IL-2 specifically in response to PDH-E2 or BCKD-E2. AD - Division of Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1984455 ID - 221 ER - TY - JOUR AU - Teuber, S. S. AU - Coppel, R. L. AU - Ansari, A. A. AU - Leung, P. S. AU - Neve, R. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 1991 TI - The identification and cloning of the murine genes encoding the liver specific F alloantigens SP - 857-70 N1 - Dec JF - J Autoimmun JO - Journal of autoimmunity VL - 4 IS - 6 SN - 0896-8411 (Print) N1 - The identification and cloning of the murine genes encoding the liver specific F alloantigens N1 - 1667467 N1 - Ca 20816/ca/nci Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Amino Acid Sequence Animals Base Sequence Blotting, Western Cloning, Molecular DNA Probes Histocompatibility Antigens Class II/genetics Immunization Isoantigens/*genetics/isolation & purification Liver/*immunology Mice Mice, Inbred Strains/genetics Molecular Sequence Data Mutation Recombinant Fusion Proteins/immunology Sequence Homology, Nucleic Acid N2 - The liver specific F alloantigen is a highly conserved abundant protein found in hepatic cytoplasm; smaller amounts are detected in renal tubule cells and the perikaryon cells of the central nervous system. Although the biological function of the F alloantigen is unknown, the immune response to F has been extensively studied as a murine model of tolerance and autoimmunity. Murine F exists in two allelic forms, designated F type 1 and type 2, each of approximately 43 kDa. The immune response to the allotypic form is restricted to mouse strains of I-Ak. Responding strains immunized with allotypic F break tolerance and produce precipitating antibody that reacts with both allelic forms, i.e., immunogen and self. Thus an autoantibody is produced. Using the previously isolated rat F cDNA as a probe, we report the cloning and sequencing of the two murine F allotypes. These two alleles are nearly homologous except at the extremes of the coding sequence. There are a number of regions within the F sequence that are similar to peptides that interact specifically with I-Ak. In particular, there is a sequence near the carboxy terminus, where the two allotypes differ, that has homology to the I-Ak restricted malarial antigen peptide of the ring-infected erythrocyte surface antigen (RESA). AD - Division of Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1667467 ID - 216 ER - TY - JOUR AU - Smythe, J. A. AU - Coppel, R. L. AU - Day, K. P. AU - Martin, R. K. AU - Oduola, A. M. AU - Kemp, D. J. AU - Anders, R. F. PY - 1991 TI - Structural diversity in the Plasmodium falciparum merozoite surface antigen 2 SP - 1751-5 N1 - Mar 1 JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 88 IS - 5 SN - 0027-8424 (Print) N1 - Structural diversity in the Plasmodium falciparum merozoite surface antigen 2 N1 - 2000383 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Antigens, Surface/*genetics Base Sequence Genes Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotide Probes Plasmodium falciparum/genetics/*immunology Polymerase Chain Reaction Sequence Homology, Nucleic Acid *Variation (Genetics) N2 - Antigens associated with the surface of merozoites of the malaria parasite Plasmodium falciparum are directly accessible to immune attack and therefore are prime vaccine candidates. We have previously shown that one of the two known merozoite surface antigens (merozoite surface antigen 2; MSA-2) exhibits considerable sequence and antigenic diversity in different isolates. The sequences of MSA-2 from three isolates revealed a central domain composed of repeats that vary in number, length, and sequence, flanked in turn by nonrepetitive variable sequences and by conserved N- and C-terminal domains. We report here the sequences of a further four MSA-2 alleles, containing repetitive sequences that are related but not identical to each other. The seven alleles of MSA-2 can be divided into two distinct allele families on the basis of nonrepetitive sequences. Hybridization studies with repeat probes indicated that all of the 44 P. falciparum isolates examined contained repeat regions similar to those defined in known MSA-2 sequences. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2000383 ID - 220 ER - TY - JOUR AU - Marshall, V. M. AU - Coppel, R. L. AU - Martin, R. K. AU - Oduola, A. M. AU - Anders, R. F. AU - Kemp, D. J. PY - 1991 TI - A Plasmodium falciparum MSA-2 gene apparently generated by intragenic recombination between the two allelic families SP - 349-51 N1 - Apr JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 45 IS - 2 SN - 0166-6851 (Print) N1 - A Plasmodium falciparum MSA-2 gene apparently generated by intragenic recombination between the two allelic families N1 - 2038365 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/*genetics/immunology Antigens, Surface/genetics/immunology Base Sequence DNA, Protozoan/genetics Molecular Sequence Data Plasmodium falciparum/*genetics/immunology Protozoan Proteins/*genetics/immunology *Recombination, Genetic Repetitive Sequences, Nucleic Acid AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2038365 ID - 219 ER - TY - JOUR AU - Leung, P. S. AU - Van de Water, J. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1991 TI - Molecular characterization of the mitochondrial autoantigens in primary biliary cirrhosis SP - 518-27 JF - Immunol Res JO - Immunologic research VL - 10 IS - 3-4 SN - 0257-277X (Print) N1 - Molecular characterization of the mitochondrial autoantigens in primary biliary cirrhosis N1 - 1720161 N1 - Dk39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. Review Switzerland N1 - eng KW - Amino Acid Sequence Autoantigens/*chemistry Epitopes/chemistry Humans Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/*immunology Molecular Sequence Data AD - Division of Rheumatology, Allergy, Clinical Immunology, University of California, Davis. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1720161 ID - 222 ER - TY - JOUR AU - Iwayama, T. AU - Leung, P. S. AU - Coppel, R. L. AU - Roche, T. E. AU - Patel, M. S. AU - Mizushima, Y. AU - Nakagawa, T. AU - Dickson, R. AU - Gershwin, M. E. PY - 1991 TI - Specific reactivity of recombinant human PDC-E1 alpha in primary biliary cirrhosis SP - 769-78 N1 - Oct JF - J Autoimmun JO - Journal of autoimmunity VL - 4 IS - 5 SN - 0896-8411 (Print) N1 - Specific reactivity of recombinant human PDC-E1 alpha in primary biliary cirrhosis N1 - 1797026 N1 - Dk 20478/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. England N1 - eng KW - Autoantibodies/biosynthesis Autoantigens Blotting, Western Cholangitis, Sclerosing/immunology Chromosome Mapping Cloning, Molecular Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Hepatitis, Chronic/immunology Humans Liver Cirrhosis, Biliary/*immunology Mitochondria/immunology Peptides/genetics/*immunology Pyruvate Dehydrogenase Complex/genetics/*immunology Recombinant Proteins/genetics/immunology Transformation, Genetic N2 - The mitochondrial autoantigens recognized by autoantibodies in patients with primary biliary cirrhosis have been identified as components of related multi-enzyme complexes, including acyltransferases of the pyruvate dehydrogenase complex (PDC), the branched-chain alpha-keto acid dehydrogenase complex (BCODH), the alpha-ketoglutarate dehydrogenase complex (OGDC), protein X and pyruvate dehydrogenase (PDC) E1 alpha and E1 beta. The major autoantigens, PDC-E2, BCODH-E2 and OGDC-E2, share some sequence homology; the epitopes on these antigens appear to be close to, or identical with, the lipoic acid binding site. Furthermore, all three antigens share some structural homology. In contrast, antibodies to PDC-E1 alpha are present in lower titers, and have been more difficult to detect. PDC-E1 alpha also differs from the three major autoantigens in that it lacks any covalently bound lipoic acid. PDC-E1 alpha cannot be purified in large quantities and becomes unstable in the absence of PDC-E1 beta. To address these problems, we have subcloned recombinant human PDC-E1 alpha to pGEX, pGEX is a vector which produces a recombinant polypeptide fused to glutathione S-transferase. The resultant E1 alpha fusion protein is stable and has a low background in immunoassays. Using the recombinant protein, we have developed an ELISA that allows rapid and reproducible quantification of antibodies to human PDC-E1 alpha. Finally, we demonstrate that a major epitope on PDC-E1 alpha is within a 300 amino acid region that contains the enzyme functional sites, namely the phosphorylation site and the TPP binding site. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1797026 ID - 218 ER - TY - JOUR AU - Brown, H. J. AU - Coppel, R. L. PY - 1991 TI - Primary structure of a Plasmodium falciparum rhoptry antigen SP - 99-110 N1 - Nov JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 49 IS - 1 SN - 0166-6851 (Print) N1 - Primary structure of a Plasmodium falciparum rhoptry antigen N1 - 1775161 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence Chromosome Mapping DNA, Protozoan/genetics Exons Molecular Sequence Data Plasmodium falciparum/*genetics/*immunology Protozoan Proteins/genetics/immunology N2 - The high-molecular-weight rhoptry complex of Plasmodium falciparum consists of 3 non-covalently associated polypeptides of 150, 135 and 105 kDa. We present the complete nucleotide sequence of the 105-kDa (RhopH3) component of this complex derived from analysis of genomic and cDNA clones. The genomic structure is unusually complex for P. falciparum, consisting of 7 exons including 2 mini-exons of 19 and 21 amino acids. The sequence lacks tandem repeats and is conserved among several parasite isolates. B cell epitopes that induce antibody responses during natural infection were mapped to five different regions of the polypeptide. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1775161 ID - 217 ER - TY - JOUR AU - Surh, C. D. AU - Coppel, R. AU - Gershwin, M. E. PY - 1990 TI - Structural requirement for autoreactivity on human pyruvate dehydrogenase-E2, the major autoantigen of primary biliary cirrhosis. Implication for a conformational autoepitope SP - 3367-74 N1 - May 1 JF - J Immunol VL - 144 IS - 9 SN - 0022-1767 (Print) N1 - Structural requirement for autoreactivity on human pyruvate dehydrogenase-E2, the major autoantigen of primary biliary cirrhosis. Implication for a conformational autoepitope N1 - 1691756 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - Amino Acid Sequence Antigens, Bacterial/immunology Autoantibodies/*immunology Autoantigens/*immunology Cross Reactions Epitopes Humans Liver Cirrhosis, Biliary/*immunology Molecular Sequence Data Molecular Weight Protein Conformation Pyruvate Dehydrogenase (Lipoamide) Pyruvate Dehydrogenase Complex/genetics/*immunology Recombinant Fusion Proteins/immunology Restriction Mapping N2 - The E2 component (acetyltransferase) of the pyruvate dehydrogenase (PDH) complex is the major mitochondrial autoantigen recognized by autoantibodies in patients with primary biliary cirrhosis (PBC). Previous work, using only a partial length rat liver cDNA clone of PDH-E2, demonstrated that the immunodominant epitope was localized to the lipoic acid binding site. Human PDH-E2, in contrast to rat PDH-E2, has two lipoic acid binding sites. By using a full length human cDNA for PDH-E2, and by preparation of multiple overlapping recombinant fragments, we have determined that three autoreactive determinants are present on human PDH-E2: two cross-reactive lipoyl domains, and an area surrounding the E1/E3 binding region. The dominant epitope was localized to the inner lipoyl domain whereas the outer lipoyl domain only showed a weak cross-reactivity, and only 1/26 PBC sera reacted weakly to the E1/E3 binding region area. By probing recombinant fusion proteins expressed from small restriction fragments of the inner lipoyl domain, we have found that a minimum of 75 amino acids (residues 146-221) were required for detectable autoantibody binding, and that 93 amino acids (residues 128-221) were necessary for characteristically strong antimitochondrial autoantibody recognition. Such a requirement for a large region suggests the possibility that a conformational autoepitope may be recognized. In addition, we have found that absorption of PBC sera with the purified mammalian PDH complex does not remove reactivity against Escherichia coli Ag. The possible implications for such results are discussed. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1691756 ID - 229 ER - TY - JOUR AU - Smythe, J. A. AU - Peterson, M. G. AU - Coppel, R. L. AU - Saul, A. J. AU - Kemp, D. J. AU - Anders, R. F. PY - 1990 TI - Structural diversity in the 45-kilodalton merozoite surface antigen of Plasmodium falciparum SP - 227-34 N1 - Mar JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 39 IS - 2 SN - 0166-6851 (Print) N1 - Structural diversity in the 45-kilodalton merozoite surface antigen of Plasmodium falciparum N1 - 2181307 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics/immunology Antigens, Surface/genetics/immunology Base Sequence Genes Molecular Sequence Data Plasmodium falciparum/genetics/*immunology Repetitive Sequences, Nucleic Acid Sequence Homology, Nucleic Acid *Variation (Genetics) N2 - An integral membrane protein associated with the merozoite surface of Plasmodium falciparum termed merozoite surface antigen 2 (the 45-kDa merozoite surface antigen), occurs in antigenically diverse forms. Here we report the sequences of the MSA 2 gene from two other isolates of P. falciparum. The 43 N-terminal residues and the 74 C-terminal residues of all three MSA 2 sequences are highly conserved, but between these conserved regions there are dramatic differences among the alleles. Instead of the two copies of a 32-amino-acid repeat present in the MSA 2 of isolate FC27, MSA 2 from clone 3D7 and isolate Indochina 1 contain 5 and 12 copies respectively of the four amino acid sequence Gly Gly Ser Ala. The sequences flanking the repeats also differ among the three antigens. The repeats in MSA 2 appear to be immunodominant during natural infection, and antibodies to the repeat regions of different alleles react with a restricted number of parasite isolates. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2181307 ID - 230 ER - TY - JOUR AU - Robertson, C. A. AU - Coppel, R. L. AU - Prindiville, T. AU - Fregeau, D. AU - Kaplan, M. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1990 TI - The relative affinity of recombinant dihydrolipoamide transacetylase for autoantibodies in primary biliary cirrhosis SP - 717-22 N1 - May JF - Hepatology JO - Hepatology (Baltimore, Md VL - 11 IS - 5 SN - 0270-9139 (Print) N1 - The relative affinity of recombinant dihydrolipoamide transacetylase for autoantibodies in primary biliary cirrhosis N1 - 2347544 N1 - Journal Article United states N1 - eng KW - Acetyltransferases/*immunology Antibody Affinity Antigen-Antibody Reactions/drug effects Autoantibodies/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Humans Liver Cirrhosis, Biliary/enzymology/*immunology *Pyruvate Dehydrogenase Complex Recombinant Proteins Thiocyanates/pharmacology N2 - In normal individuals there is an adaptive immune response to a foreign antigen in which antibodies of increasing affinity are produced with time. This is not always true of an autoimmune response. However, because only a limited number of autoantigens have been cloned or purified, this issue has not been studied well. In primary biliary cirrhosis the predominant manifestation of autoimmunity is antimitochondrial antibodies that react with dihydrolipoamide transacetylase. The availability of recombinant dihydrolipoamide transacetylase and the development of a rapid and reproducible enzyme-linked immunosorbent assay for autoantibodies has allowed us to address the affinity of autoantibodies using thiocyanate inhibition. Thiocyanate is a chaotropic compound known to inhibit antigen-antibody binding in a concentration-dependent manner. We used this property to inhibit the binding by enzyme-linked immunosorbent assay of human recombinant dihydrolipoamide transacetylase with serum autoantibodies from 55 patients with primary biliary cirrhosis. The relative affinity and serum autoantibody titers were then compared with the histological stage of the liver biopsy sample. Interestingly, we found a considerable heterogeneity of relative affinities. These relative affinities did not correlate with the histological stage or the serum titer of antimitochondrial antibodies. However, the ability of serum autoantibodies to inhibit intact primary biliary cirrhosis enzyme activity was found to correlate highly (R2 = 0.751) with the relative affinity. Thus there are profound differences between patients with respect to qualitative expression of autoantibodies. The significance of this data will be unclear until more is determined regarding the nature of the epitope that drives T cells and leads to B-cell responses. AD - Department of Internal Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2347544 ID - 228 ER - TY - JOUR AU - O'Brien, R. M. AU - Cram, D. S. AU - Coppel, R. L. AU - Harrison, L. C. PY - 1990 TI - T-cell epitopes on the 70-kDa protein of the (U1)RNP complex in autoimmune rheumatologic disorders SP - 747-57 N1 - Dec JF - J Autoimmun JO - Journal of autoimmunity VL - 3 IS - 6 SN - 0896-8411 (Print) N1 - T-cell epitopes on the 70-kDa protein of the (U1)RNP complex in autoimmune rheumatologic disorders N1 - 1708263 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Arthritis, Rheumatoid/immunology Dose-Response Relationship, Immunologic Epitopes Humans Lupus Erythematosus, Systemic/immunology Lymphocyte Activation Mixed Connective Tissue Disease/*immunology Recombinant Fusion Proteins/immunology Ribonucleoproteins/*immunology Ribonucleoproteins, Small Nuclear Sjogren's Syndrome/immunology T-Lymphocytes/*immunology N2 - High-titre IgG antibodies against the immunodominant 70-kDa protein of the (U1)ribonucleoprotein (RNP) complex are present in virtually 100% of patients with mixed connective tissue disease (MCTD), and less commonly in a variety of other autoimmune rheumatic diseases. As T-cell 'help' is assumed to be required for this potentially pathogenic form of immune response, investigations to define T-cell epitopes on the 70-kDa protein were undertaken. In prior studies we expressed the 70-kDa protein and a number of its fragments, spanning most of the molecule, as recombinant fusion proteins using the pGEX expression-vector system. These fusion proteins were used as antigens in the epitope mapping studies reported here. PBMC were isolated from patients with (U1)RNP-positive rheumatic diseases and from both normal controls and rheumatologic patients with other autoantibody reactivities, including those to Ro, La and dsDNA. Reactivity to the purified 70-kDa protein was assayed by thymidine incorporation and was evident only in anti-(U1)RNP positive patients but was not restricted to MCTD patients, being present also in patients with SLE and rheumatoid arthritis. The stimulation indices (SIs) observed were in the two- to five-fold range. Using the 70-kDa protein fragments, a T-cell stimulatory epitope was localized to the C-terminal 63 amino acids of the autoantigen. A T-cell line, derived from PBMC of a (U1)RNP positive patient with MCTD, also reacted predominantly with this C-terminal fragment but with an SI of approximately 15-fold. Thus, we have demonstrated the presence and specificity of autoreactive T lymphocytes to a defined peptide epitope in systemic rheumatic disease. AD - Burnet Clinical Research Unit, Walter and Eliza Hall Institute of Medical Research, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1708263 ID - 223 ER - TY - JOUR AU - Lustigman, S. AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. PY - 1990 TI - The mature-parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum associates with the erythrocyte membrane skeletal protein, band 4.1 SP - 261-70 N1 - Jan 15 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 38 IS - 2 SN - 0166-6851 (Print) N1 - The mature-parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum associates with the erythrocyte membrane skeletal protein, band 4.1 N1 - 2183050 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antigens, Protozoan/metabolism Antigens, Surface/metabolism *Cytoskeletal Proteins Erythrocytes/metabolism/*parasitology Humans Membrane Proteins/*blood Molecular Weight *Neuropeptides Peptide Mapping Phosphoproteins/blood Phosphorus Radioisotopes Plasmodium falciparum/*metabolism Protein Binding Protozoan Proteins/*metabolism N2 - Several proteins synthesized by mature asexual stages of Plasmodium falciparum interact with the erythrocyte membrane skeleton. One of these is the mature-parasite-infected erythrocyte surface antigen (MESA; also called PfEMP2), a phosphoprotein of 250-300 kDa, which is found on the internal face of the erythrocyte membrane. When MESA is precipitated with anti-MESA antibodies, another phosphoprotein of 80 kDa is co-precipitated. This 80-kDa phosphoprotein was identified by peptide mapping as the erythrocyte membrane component band 4.1. Thus, MESA is apparently anchored at the erythrocyte membrane through an association with band 4.1. Band 4.1 is more intensely phosphorylated in infected erythrocytes and is increased in relative molecular mass in erythrocytes infected by isolates of P. falciparum that cytoadhere. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2183050 ID - 232 ER - TY - JOUR AU - Leung, P. S. AU - Iwayama, T. AU - Coppel, R. L. AU - Gershwin, M. E. PY - 1990 TI - Site-directed mutagenesis of lysine within the immunodominant autoepitope of PDC-E2 SP - 1321-8 N1 - Dec JF - Hepatology JO - Hepatology (Baltimore, Md VL - 12 IS - 6 SN - 0270-9139 (Print) N1 - Site-directed mutagenesis of lysine within the immunodominant autoepitope of PDC-E2 N1 - 1701753 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Autoantigens/*genetics Base Sequence Binding Sites DNA/genetics Electrophoresis, Agar Gel Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Epitopes/genetics Humans Immunoblotting Liver Cirrhosis, Biliary/genetics/*immunology Lysine/*genetics Molecular Sequence Data *Mutagenesis, Site-Directed Pyruvate Dehydrogenase Complex/*genetics Thioctic Acid/*genetics N2 - The major autoantigens of PBC have been identified as the four closely related mitochondrial enzymes PDC-E2, BCKD-E2 OGDC-E2 and protein X. A major structural similarity of these enzymes is the presence of one or more lipoyl domains. The immunodominant epitope of each autoantigen has either been postulated or been demonstrated to be located within the lipoate binding region. However, it is not clear whether the binding of lipoic acid to the epitope is necessary for autoantibody recognition. To address this issue we have constructed by oligonucleotide site-directed mutatagenesis three mutants in the lipoyl domain of human PDC-E2. Because lipoic acid is covalently bound to the zeta-amino group of the lysine residue of PDC-E2, the mutants were designed to replace the lysine residue in the lipoyl domain with glutamine, a negatively charged amino acid; histidine, a positively charged amino acid; and tyrosine, an aromatic amino acid. Binding reactivity of sera from patients with PBC were analyzed by enzyme-linked immunosorbent assay, immunoblotting and specific absorption against each of the three mutants and control clones. All data were compared with parallel studies with a control recombinant clone, the liver-specific F alloantigen. We believe the recognition of the lipoyl domain is a reflection of the surface-exposed, hydrophilic and relatively mobile nature of this region of the autoantigen. Further studies on direct assay for the presence of lipoic acid will be needed to clarify these issues. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1701753 ID - 224 ER - TY - JOUR AU - Krams, S. M. AU - Van de Water, J. AU - Coppel, R. L. AU - Esquivel, C. AU - Roberts, J. AU - Ansari, A. AU - Gershwin, M. E. PY - 1990 TI - Analysis of hepatic T lymphocyte and immunoglobulin deposits in patients with primary biliary cirrhosis SP - 306-13 N1 - Aug JF - Hepatology JO - Hepatology (Baltimore, Md VL - 12 IS - 2 SN - 0270-9139 (Print) N1 - Analysis of hepatic T lymphocyte and immunoglobulin deposits in patients with primary biliary cirrhosis N1 - 2202637 N1 - Dk-39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Antibodies, Monoclonal/diagnostic use Cell Line Fluorescent Antibody Technique Fluorometry Humans Immunoglobulins/*metabolism Liver/metabolism/*pathology Liver Cirrhosis, Biliary/metabolism/*pathology Phenotype T-Lymphocytes/*pathology N2 - The histological findings in patients with primary biliary cirrhosis have been well-defined and are often used in the clinical staging of disease. However, it has only been with the development of reagents that phenotypically characterize the lymphoid infiltrate that attempts have been made to correlate pathophysiology with immune effector populations. Indeed, the inflammatory hepatic lesions in primary biliary cirrhosis have been described as containing CD4-positive and CD8-positive T cells. Less clear, however, have been the T cell receptors in these lesions. Further, the data on immunoglobulin deposits in hepatic lesions have been less well-defined; this deficit may be a result of the quality of polyspecific sera and difficulties in background. To address these issues, we have used a battery of well-defined monospecific and polyspecific reagents to phenotypically define the occurrence of lymphoid cells in the livers of patients undergoing transplantation. Furthermore, we have defined these same markers on T cell lines derived from liver, regional lymph node and peripheral blood. The predominant cell type in the mononuclear infiltrate is the CD3+, CD4+ T lymphocyte bearing the T cell receptor alpha beta. T cell lines from the same patients demonstrate similar findings. Of special importance, however, was the detection of CD20+ B cells and Ig+ cells in the lymphoid infiltrate. Indeed, we also readily demonstrated the presence of immunoglobulin on the surface of biliary epithelium. These data suggest that mechanisms involved in the pathophysiology of primary biliary cirrhosis may include both T cell and antibody mechanisms. The results also underscore the need to develop a functional, and not just a phenotypical, assay of the inflammatory infiltrate. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2202637 ID - 225 ER - TY - JOUR AU - Fregeau, D. R. AU - Roche, T. E. AU - Davis, P. A. AU - Coppel, R. AU - Gershwin, M. E. PY - 1990 TI - Primary biliary cirrhosis. Inhibition of pyruvate dehydrogenase complex activity by autoantibodies specific for E1 alpha, a non-lipoic acid containing mitochondrial enzyme SP - 1671-6 N1 - Mar 1 JF - J Immunol VL - 144 IS - 5 SN - 0022-1767 (Print) N1 - Primary biliary cirrhosis. Inhibition of pyruvate dehydrogenase complex activity by autoantibodies specific for E1 alpha, a non-lipoic acid containing mitochondrial enzyme N1 - 2106552 N1 - Dk 18320/dk/niddk Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - Antigen-Antibody Reactions Autoantibodies/*immunology Autoantigens/*immunology Blotting, Western Humans Kidney/enzymology Liver Cirrhosis, Biliary/*immunology Macromolecular Substances Mitochondria/*enzymology Mitochondria, Heart/enzymology Pyruvate Dehydrogenase Complex/*immunology Thioctic Acid/analysis N2 - Antimitochondrial antibodies (AMA) are serologically characteristic of patients with PBC. Four Ag recognized by AMA have been recently identified, including protein X and the acyltransferases of three related multienzyme complexes: the pyruvate dehydrogenase complex (PDC), the branched-chain alpha-ketoacid dehydrogenase complex, and the alpha-ketoglutarate dehydrogenase complex. Each of these enzymes contains one or more lipoyl moieties as part of a major functional site. In addition, epitope mapping has suggested that the AMA target is this lipoic acid-binding region. In this report we demonstrate that sera from patients with PBC also recognize the E1 component (pyruvate dehydrogenase, EC 1.2.4.1) of PDC. PDC-E1 is composed of alpha and beta-chains. Reactivity with the 41 kD alpha chain was detected in 80 of 120 (66%) PBC sera by immunoblotting against purified PDC-E1; 2 of 120 sera also demonstrated reactivity with the 34 kD beta chain. In contrast, sera from patients with SLE, chronic active hepatitis, or progressive sclerosing cholangitis as well as sera from healthy volunteers did not react with PDC-E1. Furthermore, affinity-purified PBC sera against PDC-E1 alpha were able to inhibit PDC enzyme activity, whereas control sera could not. PDC-E1 alpha is now the fifth mitochondrial autoantigen of PBC to be identified. Similar to the four previously identified autoantigens, AMA appear to be directed to a functional site of PDC-E1 alpha inasmuch as they are able to inhibit enzyme function. However, PDC-E1 alpha is also unique in that it is the first identified mitochondrial autoantigen which does not contain lipoic acid. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2106552 ID - 231 ER - TY - JOUR AU - Fregeau, D. R. AU - Prindiville, T. AU - Coppel, R. L. AU - Kaplan, M. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1990 TI - Inhibition of alpha-ketoglutarate dehydrogenase activity by a distinct population of autoantibodies recognizing dihydrolipoamide succinyltransferase in primary biliary cirrhosis SP - 975-81 N1 - Jun JF - Hepatology JO - Hepatology (Baltimore, Md VL - 11 IS - 6 SN - 0270-9139 (Print) N1 - Inhibition of alpha-ketoglutarate dehydrogenase activity by a distinct population of autoantibodies recognizing dihydrolipoamide succinyltransferase in primary biliary cirrhosis N1 - 2365294 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Absorption Acyltransferases/*immunology Autoantibodies/*immunology/physiology Humans Immunoblotting Immunoglobulin Isotypes/classification/immunology Ketoglutarate Dehydrogenase Complex/*antagonists & inhibitors/immunology Ketone Oxidoreductases/*antagonists & inhibitors Liver Cirrhosis, Biliary/*enzymology/immunology Mitochondria/immunology N2 - Sera from patients with primary biliary cirrhosis contain autoantibodies that recognize mitochondrial proteins. Five of the target autoantigens have now been identified as enzymes of three related multienzyme complexes: the pyruvate dehydrogenase complex, the branched chain alpha-ketoacid dehydrogenase complex and the alpha-ketoglutarate dehydrogenase complex. Each complex consists of component enzymes designated E1, E2 and E3. In this report, we confirm that primary biliary cirrhosis sera react with dihydrolipoamide succinyltransferase, the E2 component of alpha-ketoglutarate dehydrogenase complex. Seventy-three of 188 (39%) primary biliary cirrhosis sera reacted with alpha-ketoglutarate dehydrogenase complex-E2 when immunoblotted against purified alpha-ketoglutarate dehydrogenase complex; one of these sera also reacted with the E1 component. In addition, primary biliary cirrhosis sera possessing alpha-ketoglutarate dehydrogenase complex-E2 reactivity specifically inhibited enzyme function of alpha-ketoglutarate dehydrogenase complex. Enzyme activity was not affected by primary biliary cirrhosis sera that contained autoantibodies to pyruvate dehydrogenase complex-E2 and/or branched chain alpha-ketoacid dehydrogenase complex-E2, which lacked alpha-ketoglutarate dehydrogenase complex-E2 reactivity. Furthermore, affinity-purified primary biliary cirrhosis sera against alpha-ketoglutarate dehydrogenase complex-E2 inhibited only alpha-ketoglutarate dehydrogenase complex activity but did not alter enzyme activity of either pyruvate dehydrogenase complex or branched chain alpha-ketoacid dehydrogenase complex. Finally, alpha-ketoglutarate dehydrogenase complex-E2 specific affinity-purified antisera did not react on immunoblot with any component enzymes of pyruvate dehydrogenase complex or branched chain alpha-ketoacid dehydrogenase complex.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Division of Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2365294 ID - 227 ER - TY - JOUR AU - Cram, D. S. AU - Fisicaro, N. AU - Coppel, R. L. AU - Whittingham, S. AU - Harrison, L. C. PY - 1990 TI - Mapping of multiple B cell epitopes on the 70-kilodalton autoantigen of the U1 ribonucleoprotein complex SP - 630-5 N1 - Jul 15 JF - J Immunol VL - 145 IS - 2 SN - 0022-1767 (Print) N1 - Mapping of multiple B cell epitopes on the 70-kilodalton autoantigen of the U1 ribonucleoprotein complex N1 - 1694884 N1 - Journal Article Research Support, Non-U.S. Gov't United states 1950) N1 - eng KW - Autoantigens/*immunology Autoimmune Diseases/*immunology B-Lymphocytes/*immunology Base Sequence Epitopes Gene Products, gag/immunology Humans Molecular Sequence Data Molecular Weight Oligonucleotide Probes Recombinant Fusion Proteins/immunology Ribonucleoproteins/genetics/*immunology Ribonucleoproteins, Small Nuclear N2 - High titer IgG autoantibodies to the 70-kDa polypeptide component (p70) of the U1 ribonucleoprotein (RNP) complex occur in the sera of patients with mixed connective tissue disease, SLE, and related rheumatic diseases. To gain insight into the pathogenesis and diversity of this antibody response we have used recombinant DNA technology to map the linear B cell epitopes on p70. A full length 1.7-kb cDNA clone encoding p70 was isolated from a human placental library and restriction fragments or polymerase chain reaction-generated fragments of the gene subcloned into the bacterial expression vector pGEX. Purified fusion proteins representing specific regions of p70 were immunoblotted with a panel of 70 anti-(U1)RNP+ sera containing anti-p70 antibodies. Six epitopes, four major (A, B, C, and F) and two minor (D and E) were mapped and were located throughout the molecule. The anti-(U1)RNP sera displayed heterogeneity in their pattern of reactivity to the six epitopes although reactivity to epitope C was more frequently associated with SLE rather than mixed connective tissue disease. The identification of multiple B cell epitopes on p70 is consistent with the concept that this self Ag drives the autoantibody response. AD - Burnet Clinical Research Unit, Walter and Eliza Hall Institute of Medical Research, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1694884 ID - 226 ER - TY - JOUR AU - Van de Water, J. AU - Surh, C. D. AU - Leung, P. S. AU - Krams, S. M. AU - Fregeau, D. AU - Davis, P. AU - Coppel, R. AU - Mackay, I. R. AU - Gershwin, M. E. PY - 1989 TI - Molecular definitions, autoepitopes, and enzymatic activities of the mitochondrial autoantigens of primary biliary cirrhosis SP - 132-7 N1 - May JF - Semin Liver Dis JO - Seminars in liver disease VL - 9 IS - 2 SN - 0272-8087 (Print) N1 - Molecular definitions, autoepitopes, and enzymatic activities of the mitochondrial autoantigens of primary biliary cirrhosis N1 - 2471277 N1 - Journal Article Review United states N1 - eng KW - Autoantigens/analysis/immunology/*physiology *Epitopes Humans Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/enzymology/*immunology Molecular Weight AD - Department of Internal Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2471277 ID - 236 ER - TY - JOUR AU - Van de Water, J. AU - Cooper, A. AU - Surh, C. D. AU - Coppel, R. AU - Danner, D. AU - Ansari, A. AU - Dickson, R. AU - Gershwin, M. E. PY - 1989 TI - Detection of autoantibodies to recombinant mitochondrial proteins in patients with primary biliary cirrhosis SP - 1377-80 N1 - May 25 JF - N Engl J Med JO - The New England journal of medicine VL - 320 IS - 21 SN - 0028-4793 (Print) N1 - Detection of autoantibodies to recombinant mitochondrial proteins in patients with primary biliary cirrhosis N1 - 2716784 N1 - Dk-39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Autoantibodies/*analysis Autoantigens/analysis Dihydrolipoamide Dehydrogenase/*immunology Enzyme-Linked Immunosorbent Assay Humans Ketone Oxidoreductases/*immunology Liver Cirrhosis, Biliary/*immunology Mitochondria/enzymology/*immunology Multienzyme Complexes/*immunology Recombinant Proteins/immunology N2 - Primary biliary cirrhosis is characterized by the presence of autoantibodies to mitochondria with specific reactivity to proteins of 74 and 52 kilodaltons (kd). The 74-kd mitochondrial protein is the E2 component--dihydrolipoamide acetyltransferase--of the pyruvate dehydrogenase complex, and the 52-kd protein is the equivalent E2 component--dihydrolipoamide acyltransferase--of the branched-chain alpha-keto acid dehydrogenase complex. Current methods for the detection of antibodies to these proteins lack specificity or sensitivity, or they are time-consuming and not readily available. We therefore developed an enzyme-linked immunoassay to quantify specific antimitochondrial antibodies in patients with primary biliary cirrhosis. Recombinant polypeptides coding for both the 74-kd and the 52-kd mitochondrial autoantigens were used to analyze 217 coded serum samples, including samples from 93 patients with primary biliary cirrhosis and 124 controls, for reactivity by our immunoassay, immunoblotting, and immunofluorescence testing. Serum samples from 89 of the 93 patients with primary biliary cirrhosis reacted with either the pyruvate dehydrogenase-E2 or the branched-chain alpha-keto acid dehydrogenase protein. None of the 124 control samples from healthy volunteers (n = 86) or patients with primary sclerosing cholangitis (n = 38) had significant reactivity. Our results indicate that the use of recombinant, cloned autoantigens provides a simple, accurate, and rapid method of quantifying and monitoring the levels of specific mitochondrial autoantibodies in the serum of patients with primary biliary cirrhosis. AD - Division of Rheumatology/Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2716784 ID - 235 ER - TY - JOUR AU - Surh, C. D. AU - Roche, T. E. AU - Danner, D. J. AU - Ansari, A. AU - Coppel, R. L. AU - Prindiville, T. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1989 TI - Antimitochondrial autoantibodies in primary biliary cirrhosis recognize cross-reactive epitope(s) on protein X and dihydrolipoamide acetyltransferase of pyruvate dehydrogenase complex SP - 127-33 N1 - Aug JF - Hepatology JO - Hepatology (Baltimore, Md VL - 10 IS - 2 SN - 0270-9139 (Print) N1 - Antimitochondrial autoantibodies in primary biliary cirrhosis recognize cross-reactive epitope(s) on protein X and dihydrolipoamide acetyltransferase of pyruvate dehydrogenase complex N1 - 2473022 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acetyltransferases/*immunology *Antigen-Antibody Reactions Autoantibodies/*immunology Blotting, Western Cross Reactions Dihydrolipoyllysine-Residue Acetyltransferase Electrophoresis, Gel, Two-Dimensional Epitopes/*immunology Humans Liver Cirrhosis, Biliary/*immunology Mitochondria, Heart/*immunology Peptides/*immunology Pyruvate Dehydrogenase Complex/*immunology Species Specificity N2 - Antimitochondrial autoantibodies are characteristically present in sera of patients with primary biliary cirrhosis. The antimitochondrial autoantibodies recognize four major antigens from beef heart mitochondria at relative molecular weights of 74, 56, 52 and 48 kD. In the present study, we report that the 56 kD antigen is the protein X of pyruvate dehydrogenase complex and that it possesses cross-reactive antimitochondrial autoantibody epitope(s) with the 74 kD antigen, the acetyltransferase (E2) of the pyruvate dehydrogenase complex. This was demonstrated by comparing the specificities of primary biliary cirrhosis sera with a protein X-specific rabbit antiserum and by absorbing primary biliary cirrhosis sera with recombinant pyruvate dehydrogenase-E2 fusion protein. In the two-dimensional gel analysis, primary biliary cirrhosis sera and protein X-specific rabbit antiserum reacted to the same two isoelectric point polypeptides at 56 kD molecular weight. The absorption of primary biliary cirrhosis sera with the human recombinant pyruvate dehydrogenase-E2 removed reactivity toward both the 74 and 56 kD antigens. Furthermore, analysis of 82 antimitochondrial autoantibody-positive primary biliary cirrhosis sera by immunoblotting did not reveal any sera which reacted solely against either the 74 or 56 kD antigen. Finally, primary biliary cirrhosis sera recognized protein X from human, bovine and porcine sources but not protein X from rat or mouse origin. The identification of protein X as another major target of the autoimmune response in primary biliary cirrhosis suggests that the pyruvate dehydrogenase complex may have a central role in the induction of this enigmatic disease. AD - Department of Internal Medicine, School of Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2473022 ID - 233 ER - TY - JOUR AU - Surh, C. D. AU - Danner, D. J. AU - Ahmed, A. AU - Coppel, R. L. AU - Mackay, I. R. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1989 TI - Reactivity of primary biliary cirrhosis sera with a human fetal liver cDNA clone of branched-chain alpha-keto acid dehydrogenase dihydrolipoamide acyltransferase, the 52 kD mitochondrial autoantigen SP - 63-8 N1 - Jan JF - Hepatology JO - Hepatology (Baltimore, Md VL - 9 IS - 1 SN - 0270-9139 (Print) N1 - Reactivity of primary biliary cirrhosis sera with a human fetal liver cDNA clone of branched-chain alpha-keto acid dehydrogenase dihydrolipoamide acyltransferase, the 52 kD mitochondrial autoantigen N1 - 2908870 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acyltransferases/genetics/*immunology Animals Autoantigens/*genetics Cattle DNA/genetics DNA Probes Electrophoresis, Gel, Two-Dimensional Humans Isoelectric Point Liver Cirrhosis, Biliary/enzymology/genetics/*immunology Mitochondria, Liver/enzymology/*immunology Molecular Weight Multienzyme Complexes/genetics Rats Recombinant Fusion Proteins/immunology N2 - Antimitochondrial autoantibodies recognizing 68 to 74 and 50 to 52 kD inner membrane mitochondrial antigens are characteristically present in sera of patients with primary biliary cirrhosis. The biochemical identification of the antigens, however, has remained elusive. We report herein that the 52 kD antigen is the dihydrolipoamide acyltransferase of the branched-chain alpha-keto acid dehydrogenase complex. This was demonstrated by three experiments through the use of recombinant fusion protein expressed in Escherichia coli from a cDNA insert encoding the human autoantigen. First, 33 of 37 primary biliary cirrhosis patients exhibiting reactivity toward the 50 to 52 kD mitochondrial antigen by immunoblotting also showed reactivity toward the recombinant fusion protein. Second, absorption of primary biliary cirrhosis sera with recombinant fusion protein, but not with an irrelevant recombinant clone, the F-specific rat liver antigen, was effective in absorbing out reactivity against the 50 to 52 kD mitochondrial antigen but not the 68 to 74 kD antigen. Third, complete removal of reactivity toward all four different isoelectric point polypeptides at 50 to 52 kD was observed in two-dimensional gel analysis. Furthermore, primary biliary cirrhosis sera were analyzed with mitochondria from three sources, rat liver, human placenta and bovine heart, in order to compare reactivity patterns and to determine precisely the comparative molecular weights of the autoantigens in the three species. The availability of recombinant autoantigens will provide improved diagnostic tests and, more importantly, will allow definite issues in primary biliary cirrhosis to be studied, including identification of immunodominant epitopes, the significance of autoantigen recognition and the establishment of autoreactive T cell clones. AD - Department of Internal Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2908870 ID - 240 ER - TY - JOUR AU - Krams, S. M. AU - Surh, C. D. AU - Coppel, R. L. AU - Ansari, A. AU - Ruebner, B. AU - Gershwin, M. E. PY - 1989 TI - Immunization of experimental animals with dihydrolipoamide acetyltransferase, as a purified recombinant polypeptide, generates mitochondrial antibodies but not primary biliary cirrhosis SP - 411-6 N1 - Mar JF - Hepatology JO - Hepatology (Baltimore, Md VL - 9 IS - 3 SN - 0270-9139 (Print) N1 - Immunization of experimental animals with dihydrolipoamide acetyltransferase, as a purified recombinant polypeptide, generates mitochondrial antibodies but not primary biliary cirrhosis N1 - 2920998 N1 - Dk 39588/dk/niddk Journal Article Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acetyltransferases/*immunology Animals Antibodies/*immunology Blood Physiology Dihydrolipoyllysine-Residue Acetyltransferase Enzyme-Linked Immunosorbent Assay Female Guinea Pigs *Immunization Liver/physiology Liver Cirrhosis, Biliary/*immunology Liver Function Tests Macaca mulatta Mitochondria, Liver/*immunology Pyruvate Dehydrogenase Complex/antagonists & inhibitors Rabbits Rats Rats, Inbred F344 Rats, Inbred Strains Recombinant Proteins N2 - The availability of recombinant mitochondrial autoantigens may permit the experimental study of the pathophysiology of primary biliary cirrhosis. Previously, we demonstrated that high-titer antibodies to the 74 kD mitochondrial autoantigen dihydrolipoamide acetyltransferase could be generated when BALB/c mice were immunized with purified recombinant protein. Based on these data, we attempted an 8-month study to induce antibodies and liver dysfunction by immunizing AKR/J, C3H/J and CBA/HeJ mice as well as rats, guinea pigs, rabbits and rhesus monkeys with purified recombinant human dihydrolipoamide acetyltransferase. Antibodies to dihydrolipoamide acetyltransferase were readily induced and detected in all species of experimental animals with species and strain differences in the titer of the responses. Of particular interest, rabbits and guinea pigs produced antibodies which were specifically reactive with the functional site of dihydrolipoamide acetyltransferase, whereas the other strains and species produced antibodies to other epitopes on the molecule. Finally, similar to data on humans with primary biliary cirrhosis, the pyruvate dehydrogenase enzyme pathway was inhibited in the presence of immunized animal sera. These data imply that features other than simply an antibody response to mitochondrial enzymes are required for the development of primary biliary cirrhosis. Further studies will be necessary to determine the mechanisms by which mitochondrial proteins elicit an immune response. AD - Department of Internal Medicine, School of Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2920998 ID - 237 ER - TY - JOUR AU - Fregeau, D. R. AU - Davis, P. A. AU - Danner, D. J. AU - Ansari, A. AU - Coppel, R. L. AU - Dickson, E. R. AU - Gershwin, M. E. PY - 1989 TI - Antimitochondrial antibodies of primary biliary cirrhosis recognize dihydrolipoamide acyltransferase and inhibit enzyme function of the branched chain alpha-ketoacid dehydrogenase complex SP - 3815-20 N1 - Jun 1 JF - J Immunol VL - 142 IS - 11 SN - 0022-1767 (Print) N1 - Antimitochondrial antibodies of primary biliary cirrhosis recognize dihydrolipoamide acyltransferase and inhibit enzyme function of the branched chain alpha-ketoacid dehydrogenase complex N1 - 2715637 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Absorption Acetyltransferases/*immunology Autoantibodies/*physiology Collodion Dihydrolipoyllysine-Residue Acetyltransferase Enzyme-Linked Immunosorbent Assay Enzymes, Immobilized Humans Immune Sera/isolation & purification Immunoblotting Ketone Oxidoreductases/*antagonists & inhibitors/isolation & purification Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/*immunology Multienzyme Complexes/*antagonists & inhibitors/isolation & purification *Pyruvate Dehydrogenase Complex N2 - Antimitochondrial antibodies (AMA) recognizing the acetyltransferase (E2) of the pyruvate dehydrogenase (PDH) complex have been previously well-documented and the immunodominant epitope mapped. In this study, we demonstrate that sera from patients with primary biliary cirrhosis (PBC) react with another lipoic acid containing acyltransferase enzyme, namely the E2 of the branched chain alpha-ketoacid dehydrogenase (BCKD) complex. Indeed, 85/120 (71%) sera from patients with PBC reacted with BCKD-E2 by immunoblotting against purified BCKD complex. In contrast, sera from patients with chronic active hepatitis or progressive sclerosing cholangitis as well as sera from healthy volunteers did not react with any component enzymes of the BCKD complex. More importantly, BCKD enzyme activity was inhibited after incubation of the BCKD complex with either PBC sera against BCKD-E2 or with affinity purified antisera to BCKD-E2. Enzyme activity was unaltered by control sera or with PBC sera that reacted with PDH-E2 but not BCKD-E2. Furthermore, immunoblots of purified mitochondria probed with PBC sera absorbed with BCKD-E2 demonstrated that BCKD-E2 and PDH-E2 are each recognized by distinct AMA populations which do not cross-react. In addition, affinity purified PBC sera against BCKD-E2 did not react with PDH-E2 nor inhibit PDH enzyme activity, thus providing further evidence that BCKD-E2 and PDH-E2 are recognized by separate AMA. These data further suggest that the BCKD-E2 epitope recognized by AMA contains, or is close to, a functional domain of this enzyme. The availability of cDNA clones encoding BCKD-E2 and PDH-E2 will allow the study of how key metabolic enzymes may be involved in the immunology and pathology of PBC. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2715637 ID - 234 ER - TY - JOUR AU - Favaloro, J. M. AU - Marshall, V. M. AU - Crewther, P. E. AU - Coppel, R. L. AU - Kemp, D. J. AU - Anders, R. F. PY - 1989 TI - cDNA sequence predicting an octapeptide-repeat antigen of Plasmodium falciparum SP - 297-9 N1 - Jan 15 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 32 IS - 2-3 SN - 0166-6851 (Print) N1 - cDNA sequence predicting an octapeptide-repeat antigen of Plasmodium falciparum N1 - 2564637 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antigens, Protozoan/*genetics Base Sequence DNA/*genetics Plasmodium falciparum/*genetics Polymorphism, Restriction Fragment Length AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2564637 ID - 239 ER - TY - JOUR AU - Coppel, R. L. AU - Gershwin, M. E. AU - Sturgess, A. D. PY - 1989 TI - Cloned autoantigens in the study and diagnosis of autoimmune diseases SP - 27-34 N1 - Feb JF - Mol Biol Med JO - Molecular biology & medicine VL - 6 IS - 1 SN - 0735-1313 (Print) N1 - Cloned autoantigens in the study and diagnosis of autoimmune diseases N1 - 2666816 N1 - Journal Article Research Support, Non-U.S. Gov't Review England N1 - eng KW - Antibodies, Anti-Idiotypic/isolation & purification Antigens, Differentiation, T-Lymphocyte/genetics Autoantigens/*genetics Autoimmune Diseases/diagnosis/genetics/*immunology Cloning, Molecular Humans Immunity, Cellular Immunosorbent Techniques Vaccines, Synthetic/isolation & purification AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2666816 ID - 238 ER - TY - JOUR AU - Van de Water, J. AU - Gershwin, M. E. AU - Leung, P. AU - Ansari, A. AU - Coppel, R. L. PY - 1988 TI - The autoepitope of the 74-kD mitochondrial autoantigen of primary biliary cirrhosis corresponds to the functional site of dihydrolipoamide acetyltransferase SP - 1791-9 N1 - Jun 1 JF - J Exp Med JO - The Journal of experimental medicine VL - 167 IS - 6 SN - 0022-1007 (Print) N1 - The autoepitope of the 74-kD mitochondrial autoantigen of primary biliary cirrhosis corresponds to the functional site of dihydrolipoamide acetyltransferase N1 - 2455013 N1 - Dk-39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acetyltransferases/*immunology Amino Acid Sequence Autoantigens/*immunology Binding Sites Cloning, Molecular Dihydrolipoyllysine-Residue Acetyltransferase Epitopes Liver Cirrhosis, Biliary/*immunology Mitochondria/enzymology/*immunology Protein Conformation *Pyruvate Dehydrogenase Complex Solubility N2 - Autoantibodies to mitochondrial antigens are characteristic of the autoimmune liver disease primary biliary cirrhosis (PBC), but the precise antigenic determinants recognized by these antibodies have not been defined. Recently, our laboratory identified a 1,370-bp rat liver cDNA clone that coded for a polypeptide recognized specifically by sera from patients with PBC but not by sera from patients with other forms of liver disease. This recombinant protein was identified as the 74-kD M2 mitochondrial inner membrane autoantigen, now known to be dihydrolipoamide acetyltransferase. In the present study, we have identified a 603-bp fragment that codes for a polypeptide containing all of the autoreactivity of the original clone. In addition, based on hydrophobicity/hydrophilicity plots of the amino acid sequence of this polypeptide segment, several peptides were synthesized and tested for reactivity by an inhibition assay using sera from patients with PBC. One peptide, defined by the amino acids AEIETDKATIGFEVQEEGYL, absorbed serum reactivity to the protein product of the original clone. Of particular interest was the finding that this peptide contains the lipoic acid binding site KATIGF of the dihydrolipoamide acetyltransferase found in the inner mitochondrial membrane. Thus, it appears that for this autoantigen, the target of the autoantibodies corresponds to a functional site of the dihydrolipoamide acetyltransferase. AD - Department of Internal Medicine, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2455013 ID - 248 ER - TY - JOUR AU - Van de Water, J. AU - Fregeau, D. AU - Davis, P. AU - Ansari, A. AU - Danner, D. AU - Leung, P. AU - Coppel, R. AU - Gershwin, M. E. PY - 1988 TI - Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function SP - 2321-4 N1 - Oct 1 JF - J Immunol VL - 141 IS - 7 SN - 0022-1767 (Print) N1 - Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function N1 - 3049806 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - Autoantibodies/*immunology/physiology Autoantigens/immunology Autoimmune Diseases/blood/*enzymology/immunology Dihydrolipoamide Dehydrogenase/*antagonists & inhibitors/immunology/isolation & purification Humans Immunoblotting Immunosorbent Techniques Liver Cirrhosis, Biliary/blood/*enzymology/immunology N2 - Autoantibodies against mitochondria occur in the sera of patients with primary biliary cirrhosis (PBC) with characteristic reactivity to an inner membrane protein of approximately 74 kDa. To precisely define these autoantigens, we recently cloned and sequenced a rat liver cDNA (pRMIT) that encodes for all of the epitopes recognized by Ig to the 74-kDa autoantigen. In the present study we have used this recombinant probe as a tool, in addition to purified enzymes, to demonstrate by immunoblotting that the 74-kDa mitochondrial autoantigen is dihydrolipoamide acetyltransferase (EC 2.3.1.12), the core protein of the pyruvate dehydrogenase complex. Furthermore, and of particular interest, inhibition of pyruvate dehydrogenase enzyme activity was demonstrated after incubation with sera from patients with PBC but not from normal volunteers or patients with chronic active hepatitis. Such inhibition was abrogated by absorption of the PBC sera with an expressing subclone of pRMIT, designated pRMIT-603. Identification of dihydrolipoamide acetyltransferase as the target of autoimmunity in PBC provides a reagent that can be used to determine mechanisms by which this molecule is recognized. It will allow study of whether autoimmune reactivity, at the humoral or T cell level, is the basis for the pathogenesis of PBC. Additionally, such data present evidence of functional inhibition of a critical metabolic enzyme. Dihydrolipoamide acetyltransferase is well-known to mitochondrial biochemistry and, similar to identified autoantigens in other autoimmune diseases, is highly conserved in evolution. AD - Division of Rheumatology and Clinical Immunology, University of California, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3049806 ID - 244 ER - TY - JOUR AU - Surh, C. D. AU - Cooper, A. E. AU - Coppel, R. L. AU - Leung, P. AU - Ahmed, A. AU - Dickson, R. AU - Gershwin, M. E. PY - 1988 TI - The predominance of IgG3 and IgM isotype antimitochondrial autoantibodies against recombinant fused mitochondrial polypeptide in patients with primary biliary cirrhosis SP - 290-5 N1 - Mar-Apr JF - Hepatology JO - Hepatology (Baltimore, Md VL - 8 IS - 2 SN - 0270-9139 (Print) N1 - The predominance of IgG3 and IgM isotype antimitochondrial autoantibodies against recombinant fused mitochondrial polypeptide in patients with primary biliary cirrhosis N1 - 3356410 N1 - Ca 20816/ca/nci Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Autoantibodies/*analysis Enzyme-Linked Immunosorbent Assay/standards Humans Immunoglobulin G/immunology Immunoglobulin Isotypes/*immunology Immunoglobulin M/immunology Liver Cirrhosis, Biliary/*immunology Mitochondria/*immunology/metabolism Molecular Weight Peptides/*immunology/metabolism Recombinant Proteins N2 - Autoantibodies against inner mitochondrial membrane proteins are a hallmark of primary biliary cirrhosis. Specifically, these antimitochondrial autoantibodies recognize two polypeptides of approximately 70 and 52 kD, respectively. Although the specificity of antimitochondrial autoantibodies has been studied for the past 2 decades, the complementary DNA encoding the major primary biliary cirrhosis-specific 70 kD antigen has only recently been cloned. The availability of the recombinant autoantigen has resulted in the development of a highly sensitive and specific ELISA to detect antimitochondrial autoantibodies and to determine their immunoglobulin isotypes. We report herein that IgG3 is the predominant isotype of antimitochondrial autoantibodies in a group of 74 primary biliary cirrhosis patients. This finding is significant in light of the genomic immunoglobulin in heavy chain gene arrangement. Ninety-three per cent of primary biliary cirrhosis patients possessed IgG3 antimitochondrial autoantibodies with titers of 1:10(3) or higher; 32% of these patients possessed titers of 10(4), 29% at 10(5) and 7% at 10(6). IgM antimitochondrial autoantibodies were next most prevalent; 63% of the patients were positive and 50% of these patients showed titers of 10(3), 43% at 10(4) and 6% at 10(5). Other isotypes were present but in much lower titer and occurrence. Isotypes of antimitochondrial autoantibodies reactive to the 52 kD antigen were also determined using immunoblotting techniques. The predominance of IgG3 and IgM were similarly observed. Finally, the serum immunoglobulin isotype levels of primary biliary cirrhosis patients were compared with healthy normal adults by radial immunodiffusion. Serum IgG3 and IgM were very elevated in primary biliary cirrhosis; with IgG3 at 5.5-fold and IgM at 4.3-fold above normals.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Department of Internal Medicine, University of California School of Medicine, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3356410 ID - 250 ER - TY - JOUR AU - Sturgess, A. D. AU - Peterson, M. G. AU - McNeilage, L. J. AU - Whittingham, S. AU - Coppel, R. L. PY - 1988 TI - Characteristics and epitope mapping of a cloned human autoantigen La SP - 3212-8 N1 - May 1 JF - J Immunol VL - 140 IS - 9 SN - 0022-1767 (Print) N1 - Characteristics and epitope mapping of a cloned human autoantigen La N1 - 2452201 N1 - Journal Article Research Support, Non-U.S. Gov't United states 1950) N1 - eng KW - Amino Acid Sequence Autoantigens/*genetics/immunology Base Sequence Cloning, Molecular DNA/genetics Epitopes Humans Molecular Sequence Data RNA, Messenger/genetics *Ribonucleoproteins N2 - The La (SS-B) polypeptide is a ribonucleoprotein against which high titer antinuclear antibodies (ANA) react in the human autoimmune disease primary Sjogren's syndrome. To identify the autoepitopes with which the ANA anti-La (anti-SS-B) reacts, we isolated a 1.4-kb cDNA clone for La from a lambda gt10 library made from a human Burkitt's cell line. This clone contained an open reading frame of 1065 bp, encoding a 40.1-kDa polypeptide that corresponded to the carboxyl-terminal end of the La protein. The predicted polypeptide sequence of the recombinant protein was highly charged and unrelated to any previously published sequence. We also compared this clone to a previously published cDNA sequence for La and demonstrated significant differences, particularly that the open reading frame in our cDNA continued for 926 additional bases 3' to a putative termination codon in the previously reported sequence. The recombinant La protein was expressed in Escherichia coli and tested for reactivity with 200 sera containing ANA of various specificities. Only the sera containing anti-La antibodies reacted with the cloned La. By expressing subclones of the La cDNA as fusion proteins with beta-galactosidase, we have localized at least one epitope for the binding of anti-La antibodies to the carboxyl-terminal 103 amino acids of the La protein. No anti-La binding could be demonstrated to the region of the La protein that had previously been predicted to contain an autoepitope for the binding of anti-La (SS-B) antibodies. Studies of cloned autoepitopes could provide important clues to the role ANA play in disease and lead to targeted intervention in the treatment of primary Sjogren's syndrome. AD - Clinical Research Unit, Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Parkville, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2452201 ID - 249 ER - TY - JOUR AU - Smythe, J. A. AU - Coppel, R. L. AU - Brown, G. V. AU - Ramasamy, R. AU - Kemp, D. J. AU - Anders, R. F. PY - 1988 TI - Identification of two integral membrane proteins of Plasmodium falciparum SP - 5195-9 N1 - Jul JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 85 IS - 14 SN - 0027-8424 (Print) N1 - Identification of two integral membrane proteins of Plasmodium falciparum N1 - 3293051 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan/isolation & purification Antigens, Protozoan/*analysis/genetics Base Sequence Cloning, Molecular DNA/genetics/isolation & purification DNA, Recombinant/isolation & purification Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Fluorescent Antibody Technique Humans Immunoassay Immunosorbent Techniques *Membrane Glycoproteins Membrane Proteins/*analysis/genetics/immunology Molecular Sequence Data Molecular Weight Phosphatidylinositols/immunology Plasmodium falciparum/*immunology *Protozoan Proteins Sequence Homology, Nucleic Acid N2 - We describe the isolation and cloning of two integral membrane protein antigens of Plasmodium falciparum. The antigens were isolated by Triton X-114 temperature-dependent phase separation, electrophoretically transferred to nitrocellulose, and used to affinity-purify monospecific human antibodies. These antibodies were used to isolate the corresponding cDNA clones from a phage lambda gt11-Amp3 cDNA expression library. Clone Ag512 corresponds to a Mr 55,000 merozoite rhoptry antigen, and clone Ag513 corresponds to a Mr 45,000 merozoite surface antigen. Both proteins can be biosynthetically labeled with [3H]glucosamine and [3H]myristic acid, suggesting that they may be anchored in membranes via a glycosylphosphatidylinositol moiety. Similarities in the C-terminal sequences of the Mr 45,000 merozoite surface antigen and the Trypanosoma brucei variant surface glycoproteins provides further evidence that this antigen has a glycosylphosphatidylinositol anchor. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3293051 ID - 246 ER - TY - JOUR AU - Peterson, M. G. AU - Crewther, P. E. AU - Thompson, J. K. AU - Corcoran, L. M. AU - Coppel, R. L. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1988 TI - A second antigenic heat shock protein of Plasmodium falciparum SP - 71-8 N1 - Mar JF - DNA JO - DNA (Mary Ann Liebert, Inc VL - 7 IS - 2 SN - 0198-0238 (Print) N1 - A second antigenic heat shock protein of Plasmodium falciparum N1 - 3282854 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antibodies/isolation & purification Antigens, Protozoan/*genetics/immunology *Base Sequence Cloning, Molecular Electrophoresis, Polyacrylamide Gel Heat-Shock Proteins/*genetics/immunology Immunochemistry Molecular Sequence Data Plasmodium falciparum/*genetics/immunology *Sequence Homology, Nucleic Acid N2 - We describe here an antigen of Plasmodium falciparum, defined by a cDNA clone designated Ag361. The antigen is a soluble cytoplasmic 70-kD polypeptide present in all isolates analyzed and in all stages of asexual development in the blood. The antigen is a natural immunogen, although it lacks repeating epitopes of many P. falciparum antigens. Ag361 shares extensive sequence homology with the hsp70 proteins of Xenopus laevis, Drosophila melanogaster, Escherichia coli, and man, as well as a previously isolated P. falciparum hsp70 protein. The genome of P. falciparum contains at least five hsp70-like genes, located on at lest four different chromosomes. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3282854 ID - 251 ER - TY - JOUR AU - Peterson, M. G. AU - Coppel, R. L. AU - Moloney, M. B. AU - Kemp, D. J. PY - 1988 TI - Third form of the precursor to the major merozoite surface antigens of Plasmodium falciparum SP - 2664-7 N1 - Jun JF - Mol Cell Biol JO - Molecular and cellular biology VL - 8 IS - 6 SN - 0270-7306 (Print) N1 - Third form of the precursor to the major merozoite surface antigens of Plasmodium falciparum N1 - 3043189 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Alleles Animals Antigens, Protozoan/*genetics Base Sequence DNA/metabolism Molecular Sequence Data Nucleic Acid Hybridization Plasmodium falciparum/*genetics/immunology Protein Precursors/*genetics N2 - The precursor to the major merozoite surface antigens of Plasmodium falciparum appears to be encoded by two distinctly different (dimorphic) alleles able to undergo limited recombination. We analyzed 18 previously uncharacterized P. falciparum isolates to test the dimorphic model. All but one, a Thailand isolate, conformed to the dimorphic model, and this isolate conformed to the dimorphic model in all but variable block 2. Sequence analysis revealed that block 2 of isolate CSL2 was a third form. Hence, the dimorphic model is not strictly correct. Recombination between alleles was found only within two conserved blocks near the 5' end of the gene. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3043189 ID - 247 ER - TY - JOUR AU - Peterson, M. G. AU - Coppel, R. L. AU - McIntyre, P. AU - Langford, C. J. AU - Woodrow, G. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1988 TI - Variation in the precursor to the major merozoite surface antigens of Plasmodium falciparum SP - 291-301 N1 - Jan 15 JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 27 IS - 2-3 SN - 0166-6851 (Print) N1 - Variation in the precursor to the major merozoite surface antigens of Plasmodium falciparum N1 - 2449612 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/*genetics/immunology Antigens, Surface/*genetics/immunology Base Sequence Crossing Over, Genetic DNA/genetics Epitopes/immunology Genes Molecular Sequence Data Plasmodium falciparum/genetics/*immunology Protein Precursors/genetics/immunology Variation (Genetics) N2 - The major antigens on the surface of Plasmodium falciparum merozoites are derived from a single high molecular weight polypeptide. The precursor to the major merozoite surface antigen (PMMSA) has conserved and variable antigenic determinants and varies in size in different isolates. Since the protein is a candidate for a malaria vaccine, it is important to understand the molecular basis of this variation. We present the complete sequence of the PMMSA of the Papua New Guinea isolate FC27 and the partial sequence of the West African isolate NF7. The gene consists of blocks of sequence which are either conserved or variable between different isolates. Variable sequences fall into two distinct types, indicating that the PMMSA is encoded by dimorphic alleles that undergo recombination within conserved blocks at the 5' end of the gene. Genetic exchange is not apparent within the other two-thirds of the gene in 12 isolates, suggesting strong selection against such recombinants. The most variable block located near the 5' end contains repeats that are different in independent cloned isolates. This variation presumably accounts for much of the size and antigenic variability. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2449612 ID - 255 ER - TY - JOUR AU - Mu, F. T. AU - Alderuccio, F. AU - Toh, B. H. AU - Gershwin, M. E. AU - Coppel, R. AU - Mackay, I. R. PY - 1988 TI - Murine monoclonal antibody to mitochondria reacts with the 72 kD antigen of primary biliary cirrhosis SP - 100-6 N1 - Jan JF - Clin Exp Immunol JO - Clinical and experimental immunology VL - 71 IS - 1 SN - 0009-9104 (Print) N1 - Murine monoclonal antibody to mitochondria reacts with the 72 kD antigen of primary biliary cirrhosis N1 - 3280176 N1 - Journal Article England N1 - eng KW - Animals Antibodies, Monoclonal/diagnostic use Autoantigens/*analysis Dogs Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Humans Isoelectric Focusing Liver Cirrhosis, Biliary/*immunology Mice Mice, Inbred BALB C Mitochondria/*immunology Rats Swine N2 - BALB/c mice were immunized with canine gastric mucosal cells enriched to 70% for parietal cells, to produce monoclonal antibodies (MoAb). Three MoAb, FMM-4C5, FMM-4C9 and FMM-2B2, were obtained which reacted by indirect immunofluorescence with gastric parietal cells and kidney tubules, predominantly distal kidney tubules, with a pattern similar to that of the M2 autoantibodies of primary biliary cirrhosis (PBC). The antibodies also reacted with tissues from rabbit, rat, pig, human and with rod-shaped structures in acetone-fixed monolayer cultures of human fibroblasts and HEp 2 cells. FMM-4C9 and FMM-2B2 reacted with tissues from BALB/c mice but FMM-4C5 did not. Immunoblots of FMM-4C5 with mitochondrial fractions showed that the antibody recognized a 63 kD antigen from dog stomach, rat kidney and rat liver, and a 72 kD antigen from human placenta; mouse preparations were not reactive. The antigen co-migrated with that recognized by serum from cases of PBC and some cases of progressive systemic sclerosis. Absorption of the mitochondrial fraction with PBC sera removed reactivity by immunoblotting with the murine autoantibody and vice versa. Two dimension immunoblots showed that the murine and human antibodies recognized an identical series of paired 'spots'. FMM-4C5 also reacted by immunoblotting with a rat recombinant mitochondrial polypeptide which has disease-specific reactivity with PBC sera. Absorption with recombinant polypeptide removed anti-mitochondrial activity by immunoblotting and immunofluorescence. These observations suggest that the MoAb FMM-4C5 recognizes part of the same 72 kD molecule recognized by human PBC sera. The murine monoclonal antibodies should be useful probes for further studies of the structure, function and possible pathogenicity of the 72 kD autoantigen. AD - Department of Pathology and Immunology, Monash University Medical School, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3280176 ID - 256 ER - TY - JOUR AU - Lustigman, S. AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. PY - 1988 TI - A component of an antigenic rhoptry complex of Plasmodium falciparum is modified after merozoite invasion SP - 217-24 N1 - Sep JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 30 IS - 3 SN - 0166-6851 (Print) N1 - A component of an antigenic rhoptry complex of Plasmodium falciparum is modified after merozoite invasion N1 - 3054533 N1 - In Vitro Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antigens, Protozoan/*immunology/isolation & purification/metabolism Cloning, Molecular Electrophoresis, Polyacrylamide Gel Humans Immunoblotting Methionine/diagnostic use Organelles/*immunology/metabolism Peptide Mapping Plasmodium falciparum/cytology/*immunology Precipitin Tests Recombinant Proteins/immunology N2 - Human antibodies affinity purified on an adsorbent prepared from a cDNA clone (Ag44) expressing a portion of a rhoptry antigen were used to characterize the synthesis and fate of the antigen in the asexual blood stages of Plasmodium falciparum. The rhoptry antigen is synthesized in the mature trophozoite-stage parasites as a 103 kDa polypeptide, is present in the schizonts and merozoites as a 105 kDa polypeptide, is discharged from the rhoptries and found in the newly invaded red cells as a 110 kDa polypeptide. Anti-Ag44 antibodies immunoprecipitate the antigen and two additional polypeptides of 135 and 150 kDa from lysates of infected cells and from culture supernatants. The three polypeptides are associated in a non-covalent complex that persists in the newly invaded red cells. All the components of the high molecular weight rhoptry complex are antigenic and can be precipitated with immune human serum. The 135 kDa polypeptide is identical to a 140 kDa rhoptry antigen previously identified by a monoclonal antibody. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3054533 ID - 245 ER - TY - JOUR AU - Leung, P. S. AU - Gershwin, M. E. AU - Coppel, R. AU - Halpern, G. AU - Novey, H. AU - Castles, J. J. PY - 1988 TI - Localization, molecular weight and immunoglobulin subclass response to Aspergillus fumigatus allergens in acute bronchopulmonary aspergillosis SP - 416-21 JF - Int Arch Allergy Appl Immunol JO - International archives of allergy and applied immunology VL - 85 IS - 4 SN - 0020-5915 (Print) N1 - Localization, molecular weight and immunoglobulin subclass response to Aspergillus fumigatus allergens in acute bronchopulmonary aspergillosis N1 - 2451644 N1 - Nci 20816/ci/cid Journal Article Research Support, U.S. Gov't, P.H.S. Switzerland N1 - eng KW - Acute Disease Allergens/*analysis Antibodies, Fungal/*immunology Antigens, Fungal/analysis Aspergillosis, Allergic Bronchopulmonary/*immunology Aspergillus fumigatus/immunology Electrophoresis, Polyacrylamide Gel Epitopes Humans Immunoglobulins/classification Molecular Weight Sodium Dodecyl Sulfate N2 - A specimen of Aspergillus fumigatus was isolated from a patient with acute bronchopulmonary aspergillosis (ABPA). Cultures were allowed to propagate and were separated into spores and mycelium to greater than 95% homogeneity. Extracts of both the spores and mycelium were prepared and used for study by both dot blot analysis and immunoblotting to study the major allergens. By dot blot analysis, 22 of 22 patients with ABPA and none of 10 healthy controls reacted with mycelium when probed for both IgG and IgE anti-Aspergillus reactivity. Similar results were obtained when these extracts were separated on polyacrylamide gel electrophoresis and then probed; this included both IgG and IgE reactivity. Further, the reactivity to preparations of whole extract was absorbed with mycelium but not spores. Several distinct protein bands were detected with IgG, ranging from 30 to 110 kilodaltons (kD). In contrast, the majority of patients with ABPA, when studied for IgE reactivity, reacted only with a 70-kD protein, although 1 out of 22 patients reacted with a 30-kD protein. Subclass analysis of IgG reactivity demonstrated that 90% of reactive sera contained IgG2 and 73% IgG4 reactivity. IgG1 and IgG3 reactivity were present but less frequently detected. The use of immunoblotting will enable the identification of relevant allergens. Further, the demonstration that the allergenicity is located primarily in mycelium will allow more definitive studies of allergen isolation for studies of reactivity and, ultimately, cloning of the 70-kD protein and, finally, epitope mapping. AD - Division of Rheumatology and Allergy, University of California School of Medicine, Davis. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2451644 ID - 259 ER - TY - JOUR AU - Gershwin, M. E. AU - Coppel, R. L. AU - Mackay, I. R. PY - 1988 TI - Primary biliary cirrhosis and mitochondrial autoantigens--insights from molecular biology SP - 147-51 N1 - Jan-Feb JF - Hepatology JO - Hepatology (Baltimore, Md VL - 8 IS - 1 SN - 0270-9139 (Print) N1 - Primary biliary cirrhosis and mitochondrial autoantigens--insights from molecular biology N1 - 3276585 N1 - Journal Article Review United states N1 - eng KW - Animals Autoantigens/*immunology Autoimmune Diseases/*immunology Dihydrolipoyllysine-Residue Acetyltransferase Humans Liver Cirrhosis, Biliary/*immunology Mitochondria, Liver/immunology Mitochondrial Proteins AD - Division of Rheumatology and Clinical Immunology, University of California at Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3276585 ID - 257 ER - TY - JOUR AU - Fregeau, D. R. AU - Leung, P. S. AU - Coppel, R. L. AU - McNeilage, L. J. AU - Medsger, T. A., Jr. AU - Gershwin, M. E. PY - 1988 TI - Autoantibodies to mitochondria in systemic sclerosis. Frequency and characterization using recombinant cloned autoantigen SP - 386-92 N1 - Mar JF - Arthritis Rheum JO - Arthritis and rheumatism VL - 31 IS - 3 SN - 0004-3591 (Print) N1 - Autoantibodies to mitochondria in systemic sclerosis. Frequency and characterization using recombinant cloned autoantigen N1 - 3282519 N1 - Nc1-20816/phs Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Antibody Specificity Autoantibodies/*immunology Enzyme-Linked Immunosorbent Assay Humans Immunologic Techniques Mitochondria/*immunology Scleroderma, Systemic/*immunology N2 - Mitochondrial autoantibodies, a hallmark of primary biliary cirrhosis (PBC), have been widely described for many years in patients with systemic sclerosis, and there have been several reports of the concurrence of systemic sclerosis and PBC. However, there is very little information with respect to the significance of these autoantibodies or any definitive evidence that the antigens involved represent the mitochondrial autoantigens (M2 complex) described in PBC. We have cloned and sequenced a rat complementary DNA which encodes for all the epitopes recognized by autoantibodies to the major, or 70-kd, mitochondrial autoantigen in patients with PBC. Using this recombinant fused autoantigen, as well as by immunoblotting with human placental mitochondria, we tested for antimitochondrial antibody specificity in sera from 250 patients with systemic sclerosis. Nineteen sera (7.6%), including those from patients with CREST (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasias) and diffuse scleroderma, had reactivity with human placental mitochondria proteins by immunoblot testing. All 19 sera reacted with the M2 complex. All sera that reacted with the 70-kd protein likewise reacted with the recombinant cloned autoantigen. The predominant autoantibody isotype to the 70-kd protein was IgG3. Interestingly, the 70-kd protein is 11% proline, an amino acid which is frequently preceded by hydrophobic amino acids. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California School of Medicine, Davis 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3282519 ID - 252 ER - TY - JOUR AU - Coppel, R. L. AU - McNeilage, L. J. AU - Surh, C. D. AU - Van de Water, J. AU - Spithill, T. W. AU - Whittingham, S. AU - Gershwin, M. E. PY - 1988 TI - Primary structure of the human M2 mitochondrial autoantigen of primary biliary cirrhosis: dihydrolipoamide acetyltransferase SP - 7317-21 N1 - Oct JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 85 IS - 19 SN - 0027-8424 (Print) N1 - Primary structure of the human M2 mitochondrial autoantigen of primary biliary cirrhosis: dihydrolipoamide acetyltransferase N1 - 3174635 N1 - Dk 39588/dk/niddk Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states N1 - eng KW - Acetyltransferases/*analysis/genetics Amino Acid Sequence Animals Base Sequence DNA/analysis Dihydrolipoyllysine-Residue Acetyltransferase Humans Liver Cirrhosis, Biliary/*enzymology Molecular Sequence Data Molecular Weight *Pyruvate Dehydrogenase Complex Rats N2 - Primary biliary cirrhosis is a chronic, destructive autoimmune liver disease of humans. Patient sera are characterized by a high frequency (greater than 95%) of autoantibodies to a Mr 70,000 mitochondrial antigen, a component of the M2 antigen complex. We have identified a human cDNA clone encoding the complete amino acid sequence of this autoantigen. The predicted structure has significant similarity with the dihydrolipoamide acetyltransferase (EC 2.3.1.12) of the Escherichia coli pyruvate dehydrogenase multienzyme complex. The human sequence preserves the Glu-Thr-Asp-Lys-Ala motif of the lipoyl-binding site and has two potential binding sites. Expressed fragments of the cDNA react strongly with sera from patients with primary biliary cirrhosis but not with sera from patients with autoimmune chronic active hepatitis or sera from healthy subjects. AD - Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3174635 ID - 243 ER - TY - JOUR AU - Coppel, R. L. AU - Lustigman, S. AU - Murray, L. AU - Anders, R. F. PY - 1988 TI - MESA is a Plasmodium falciparum phosphoprotein associated with the erythrocyte membrane skeleton SP - 223-31 N1 - Dec JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 31 IS - 3 SN - 0166-6851 (Print) N1 - MESA is a Plasmodium falciparum phosphoprotein associated with the erythrocyte membrane skeleton N1 - 3065643 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antibodies, Protozoan/analysis Antigens, Protozoan/*biosynthesis Antigens, Surface/*biosynthesis Erythrocyte Membrane/*immunology Immunoassay Phosphoproteins/*biosynthesis Phosphorylation Plasmodium falciparum/*immunology Rabbits N2 - The mature-parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum is an antigenically variable, high molecular weight protein of trophozoites and schizonts that is located at the erythrocyte surface membrane. It is first synthesized at the late ring stage and continues to be synthesized until late schizogony. MESA cannot be detected on the external surface of erythrocytes infected by trophozoites and early schizonts but is located at the internal surface in association with the erythrocyte membrane skeleton. The degree of association with the membrane skeleton varies among parasite lines, being greater in knobby parasite lines. MESA is phosphorylated and is present in a similar location to another phosphoprotein, the ring-infected erythrocyte surface antigen (RESA). However, it differs from RESA in being detected at a later stage of asexual development of the parasite. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3065643 ID - 242 ER - TY - JOUR AU - Coppel, R. L. AU - Crewther, P. E. AU - Culvenor, J. G. AU - Perrin, L. H. AU - Brown, G. V. AU - Kemp, D. J. AU - Anders, R. F. PY - 1988 TI - Variation in p126, a parasitophorous vacuole antigen of Plasmodium falciparum SP - 155-66 N1 - Dec JF - Mol Biol Med JO - Molecular biology & medicine VL - 5 IS - 3 SN - 0735-1313 (Print) N1 - Variation in p126, a parasitophorous vacuole antigen of Plasmodium falciparum N1 - 3072468 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan Antigens, Protozoan/*genetics Base Sequence Cloning, Molecular DNA/isolation & purification Genes Humans Molecular Sequence Data Plasmodium falciparum/*immunology Variation (Genetics) N2 - We describe a cDNA clone expressing part of p126, a parasitophorous vacuole antigen of Plasmodium falciparum that is processed to smaller fragments about the time of schizont rupture. The amino acid sequence includes determinants that are antigenic during the course of infection. The sequence contains a string of serine residues but no other repetitive elements. Comparison of the sequence to a previously published sequence demonstrates conserved and variable sequence elements. The genomic context of the single copy gene is conserved in six different isolates. p126 is shown to be identical to Pf140, an antigen previously demonstrated to partially protect Saimiri monkeys against P. falciparum infection. AD - Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3072468 ID - 241 ER - TY - JOUR AU - Collins, W. E. AU - Pappaioanou, M. AU - Anders, R. F. AU - Campbell, G. H. AU - Brown, G. V. AU - Kemp, D. J. AU - Broderson, J. R. AU - Coppel, R. L. AU - Skinner, J. C. AU - Procell, P. M. AU - et al. PY - 1988 TI - Immunization trials with the ring-infected erythrocyte surface antigen of Plasmodium falciparum in owl monkeys (Aotus vociferans) SP - 268-82 N1 - Mar JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 38 IS - 2 SN - 0002-9637 (Print) N1 - Immunization trials with the ring-infected erythrocyte surface antigen of Plasmodium falciparum in owl monkeys (Aotus vociferans) N1 - 3281493 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antibodies, Protozoan/analysis Antigens, Protozoan/*immunology Antigens, Surface/*immunology Aotus trivirgatus Freund's Adjuvant *Immunization Malaria/immunology/*prevention & control Plasmodium falciparum/*immunology *Protozoan Proteins Vaccines/*immunology Vaccines, Synthetic/immunology N2 - A protocol was developed for the testing of blood stage vaccines against Plasmodium falciparum using Peruvian Aotus vociferans and the Indochina I/CDC strain of the parasite. Three different fused polypeptide vaccines containing elements of the ring-infected erythrocyte surface antigen molecule combined with Freund's complete and Freund's incomplete adjuvants were tested to determine their ability to protect against overwhelming infection following challenge with this highly virulent strain of P. falciparum, and to invoke antibody responses as measured by a standard indirect immunofluorescence technique. Nine of 14 immunized animals exhibited some protection. Presented are the test procedures developed for the conduct of such trials with New World monkeys and the analysis of results that led to the identification of variables selected for study in future trials. AD - Division of Parasitic Diseases, Center for Infectious Diseases, Atlanta, Georgia 30333. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3281493 ID - 253 ER - TY - JOUR AU - Bianco, A. E. AU - Crewther, P. E. AU - Coppel, R. L. AU - Stahl, H. D. AU - Kemp, D. J. AU - Anders, R. F. AU - Brown, G. V. PY - 1988 TI - Patterns of antigen expression in asexual blood stages and gametocytes of Plasmodium falciparum SP - 258-67 N1 - Mar JF - Am J Trop Med Hyg JO - The American journal of tropical medicine and hygiene VL - 38 IS - 2 SN - 0002-9637 (Print) N1 - Patterns of antigen expression in asexual blood stages and gametocytes of Plasmodium falciparum N1 - 3281492 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens, Protozoan/*analysis Antigens, Surface/analysis Erythrocytes/*parasitology Fluorescent Antibody Technique Humans Plasmodium falciparum/growth & development/*immunology *Protozoan Proteins N2 - The stage specificity and localization of 12 Plasmodium falciparum antigens were determined by immunofluorescence using acetone-fixed parasites reacted with monospecific antibodies against cloned antigens. Antibodies were prepared by immunization of rabbits with recombinant proteins or by affinity purification of human plasma against cloned antigen adsorbents. Most of the antigens occurred predominantly in mature asexual parasites, two were abundant in ring stages and three were absent in rings. Four of the 12 antigens were detected in asexual stages but not in gametocytes. Grouping of antigens by localization within blood stages was difficult because of the complexity of fluorescence patterns observed. With some antibodies, fluorescence was apparently distributed evenly over the parasites, but in other cases label was concentrated within discrete compartments or organelles. Extraparasitic intraerythrocytic fluorescence was also observed. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3281492 ID - 254 ER - TY - JOUR AU - Anders, R. F. AU - Coppel, R. L. AU - Brown, G. V. AU - Kemp, D. J. PY - 1988 TI - Antigens with repeated amino acid sequences from the asexual blood stages of Plasmodium falciparum SP - 148-72 JF - Prog Allergy JO - Progress in allergy VL - 41 SN - 0079-6034 (Print) N1 - Antigens with repeated amino acid sequences from the asexual blood stages of Plasmodium falciparum N1 - 2457214 N1 - Journal Article Research Support, Non-U.S. Gov't Review Switzerland N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan/biosynthesis Antigens, Protozoan/analysis/*immunology Cross Reactions Epitopes/analysis Humans Malaria/immunology Plasmodium falciparum/growth & development/*immunology UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2457214 ID - 258 ER - TY - JOUR AU - Whittingham, S. AU - Naselli, G. AU - McNeilage, L. J. AU - Coppel, R. L. AU - Sturgess, A. D. PY - 1987 TI - Serological diagnosis of primary Sjogren's syndrome by means of human recombinant La (SS-B) as nuclear antigen SP - 1-3 N1 - Jul 4 JF - Lancet JO - Lancet VL - 2 IS - 8549 SN - 0140-6736 (Print) N1 - Serological diagnosis of primary Sjogren's syndrome by means of human recombinant La (SS-B) as nuclear antigen N1 - 2885503 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Antibodies, Antinuclear/*analysis Autoantigens/*diagnostic use Enzyme-Linked Immunosorbent Assay Escherichia coli/immunology Humans Recombinant Proteins/diagnostic use *Ribonucleoproteins Sjogren's Syndrome/*diagnosis/immunology N2 - Human recombinant La nucleoprotein was purified from cultures of Escherichia coli containing a vector with a 1.4 kilobase cDNA encoding La; the nucleoprotein was used to test for antinuclear antibodies (ANA) to La. Serum samples from 260 patients with autoimmune diseases associated with ANA and 100 healthy subjects were tested by an enzyme-linked immunosorbent assay (ELISA). Samples from 47 (94%) of 50 patients with primary Sjogren's syndrome and 1 (7%) of 14 patients with secondary Sjogren's syndrome reacted with the recombinant La. No reactivity was demonstrated in 196 patients with other ANA-associated autoimmune diseases or in 100 healthy subjects. The study confirms the high correlation between ANA, anti-La, and primary Sjogren's syndrome and shows how gene cloning can provide large quantities of human autoantigens for use in highly specific and sensitive diagnostic assays. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2885503 ID - 264 ER - TY - JOUR AU - Saint, R. B. AU - Coppel, R. L. AU - Cowman, A. F. AU - Brown, G. V. AU - Shi, P. T. AU - Barzaga, N. AU - Kemp, D. J. AU - Anders, R. F. PY - 1987 TI - Changes in repeat number, sequence, and reading frame in S-antigen genes of Plasmodium falciparum SP - 2968-73 N1 - Aug JF - Mol Cell Biol JO - Molecular and cellular biology VL - 7 IS - 8 SN - 0270-7306 (Print) N1 - Changes in repeat number, sequence, and reading frame in S-antigen genes of Plasmodium falciparum N1 - 3313007 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Alleles Amino Acid Sequence Animals Antigens, Protozoan/*genetics Arrestin Base Sequence Cloning, Molecular *Genes Molecular Sequence Data Plasmodium falciparum/*genetics Repetitive Sequences, Nucleic Acid N2 - The S antigens from different isolates of Plasmodium falciparum exhibit extensive size, charge, and serological diversity. We show here that the S-antigen genes behave as multiple alleles of a single locus. The size heterogeneity results from different numbers, lengths, and/or sequences of tandem repeat units encoded within the S-antigen genes. Two genes studied here encode antigenically different S antigens but nevertheless have closely related tandem repeat sequences. We show that antigenic differences can arise because repeats are translated in different reading frames. AD - Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3313007 ID - 262 ER - TY - JOUR AU - McIntyre, P. AU - Coppel, R. L. AU - Smith, D. B. AU - Stahl, H. D. AU - Corcoran, L. M. AU - Langford, C. J. AU - Favaloro, J. M. AU - Crewther, P. E. AU - Brown, G. V. AU - Mitchell, G. F. AU - et al. PY - 1987 TI - Expression of parasite antigens in Escherichia coli SP - 59-67 N1 - Feb JF - Int J Parasitol JO - International journal for parasitology VL - 17 IS - 1 SN - 0020-7519 (Print) N1 - Expression of parasite antigens in Escherichia coli N1 - 3294643 N1 - Journal Article Review England N1 - eng KW - Animals Antigens, Helminth/*genetics Antigens, Protozoan/*genetics Bacteriophages *Cloning, Molecular DNA/genetics Escherichia coli/*genetics Gene Expression Regulation Genes Genetic Vectors Plasmids UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3294643 ID - 268 ER - TY - JOUR AU - Kemp, D. J. AU - Coppel, R. L. AU - Anders, R. F. PY - 1987 TI - Repetitive proteins and genes of malaria SP - 181-208 JF - Annu Rev Microbiol JO - Annual review of microbiology VL - 41 SN - 0066-4227 (Print) N1 - Repetitive proteins and genes of malaria N1 - 3318667 N1 - Journal Article Research Support, Non-U.S. Gov't Review United states N1 - eng KW - Amino Acid Sequence Animals Antibodies, Protozoan/biosynthesis Antigenic Variation Antigens, Protozoan/*genetics DNA/genetics Humans Malaria/*immunology Plasmodium/*genetics/immunology *Repetitive Sequences, Nucleic Acid AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3318667 ID - 269 ER - TY - JOUR AU - Gershwin, M. E. AU - Mackay, I. R. AU - Sturgess, A. AU - Coppel, R. L. PY - 1987 TI - Identification and specificity of a cDNA encoding the 70 kd mitochondrial antigen recognized in primary biliary cirrhosis SP - 3525-31 N1 - May 15 JF - J Immunol VL - 138 IS - 10 SN - 0022-1767 (Print) N1 - Identification and specificity of a cDNA encoding the 70 kd mitochondrial antigen recognized in primary biliary cirrhosis N1 - 3571977 N1 - Journal Article Research Support, Non-U.S. Gov't United states 1950) N1 - eng KW - Amino Acid Sequence Animals Autoantibodies/immunology Autoantigens/*genetics/immunology Base Sequence DNA/*genetics Dihydrolipoyllysine-Residue Acetyltransferase Humans Immunization Liver Cirrhosis, Biliary/*immunology Mice Mice, Inbred BALB C Mitochondria/*immunology Mitochondrial Proteins Rats Recombinant Fusion Proteins/immunology Species Specificity N2 - Mitochondrial autoantibodies are characteristic of the disease primary biliary cirrhosis (PBC), but the immunoreactive mitochondrial antigens have not been defined. We used a rat liver cDNA library in lambda gt 11-Amp3 to clone a 1370-base pair insert that coded for a polypeptide reactive with PBC sera. This insert was subcloned for expression into pBTA224, a plasmid vector in the same reading frame as lambda-Amp3. A positive clone, designated pRMIT, that expressed a fused polypeptide of 160 kd, was recognized by 25 of 25 sera from patients with PBC and none of 96 sera from normal persons or patients with systemic lupus erythematosus, rheumatoid arthritis, or chronic active hepatitis. This fused polypeptide was shown to correspond with the 70 kd mitochondrial autoantigen by several experiments. First, lysates of pRMIT in J101 absorbed out the 70 kd reactivity of PBC sera when probed against fractionated placental mitochondria. Second, affinity-purified antisera reactive with the fused polypeptide also reacted with the 70 kd mitochondrial antigen. Third, such affinity-purified antisera produced the characteristic anti-mitochondrial pattern of immunofluorescence on tissue sections. Finally, immunization of BALB/c mice with the fused polypeptide elicited antibodies to mitochondria. These murine antibodies reacted with the 70 kd mitochondrial protein and also produced typical mitochondrial immunofluorescence on tissue sections. The nucleotide and amino acid sequence of the recombinant protein, which encodes for approximately a 48 kd protein, showed no significant homologies with known proteins, and there were no homologies with mitochondrial genomic DNA. The availability of a recombinant form of the 70 kd mitochondrial autoantigen will allow several definitive questions to be addressed in PBC, including identification of B cell epitopes, T cell recognition, and a model of PBC in mice. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3571977 ID - 265 ER - TY - JOUR AU - Gershwin, M. E. AU - Coppel, R. L. AU - Bearer, E. AU - Peterson, M. G. AU - Sturgess, A. AU - Mackay, I. R. PY - 1987 TI - Molecular cloning of the liver-specific rat F antigen SP - 3828-33 N1 - Dec 1 JF - J Immunol VL - 139 IS - 11 SN - 0022-1767 (Print) N1 - Molecular cloning of the liver-specific rat F antigen N1 - 2445820 N1 - 20816/phs Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. United states 1950) N1 - eng KW - Amino Acid Sequence Animals Bacterial Proteins/genetics Base Sequence Cloning, Molecular DNA/genetics Epitopes/immunology Escherichia coli/genetics Isoantigens/*genetics Liver/*immunology Male Mice Mice, Inbred BALB C/genetics/immunology Mice, Inbred C3H Mice, Inbred CBA/genetics/immunology Molecular Sequence Data Rats Ribosomal Proteins/genetics Sequence Homology, Nucleic Acid N2 - F antigen is a 43-kDa widely conserved liver protein that has been intensively used in studies of immunogenicity and tolerance; two murine allotypes have been identified. Immunization of specific responder inbred strains with liver homogenates from the opposite allotype leads to precipitating antibody and cell-mediated immunity against F. The antibodies produced are autoantibodies as they react equally well with self. We have identified a cDNA clone from rat liver that reacts with alloantisera to F. The fused polypeptide produced by the clone was shown to correspond to F by several experiments. First, alloantisera to F antigen reacted with the cloned fused polypeptide, but not control recombinant clones. Second, mice immunized with the fused polypeptide generate an antibody response that reacts specifically with the 43-kDa protein of mouse liver homogenates and with highly purified F antigen. Finally, both anti-F allosera and sera from mice immunized with the fused polypeptide react with the same 43-kDa liver protein on two-dimensional immunoblots. The nucleotide and deduced amino acid sequence of the clone are presented and the sequence was found to have a significant homology with L28, an Escherichia coli ribosomal protein. The availability of recombinant F antigen will allow definitive questions to be addressed with respect to epitopes and specifically the identification of the T cell epitope which allows for autoimmune responses. AD - Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, 95616. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2445820 ID - 261 ER - TY - JOUR AU - Culvenor, J. G. AU - Langford, C. J. AU - Crewther, P. E. AU - Saint, R. B. AU - Coppel, R. L. AU - Kemp, D. J. AU - Anders, R. F. AU - Brown, G. V. PY - 1987 TI - Plasmodium falciparum: identification and localization of a knob protein antigen expressed by a cDNA clone SP - 58-67 N1 - Feb JF - Exp Parasitol JO - Experimental parasitology VL - 63 IS - 1 SN - 0014-4894 (Print) N1 - Plasmodium falciparum: identification and localization of a knob protein antigen expressed by a cDNA clone N1 - 3542549 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens, Protozoan/analysis/*genetics DNA/genetics Erythrocyte Membrane/analysis/ultrastructure Erythrocytes/parasitology/ultrastructure Humans Immunologic Techniques Microscopy, Electron Peptides/analysis/*genetics/immunology Plasmodium falciparum/analysis/*genetics/immunology/ultrastructure Protozoan Proteins N2 - Differential screening of cDNA libraries constructed from knobby and predominantly knobless Plasmodium falciparum isolates, identified the sequence SD17. Chromosome blotting experiments have shown that this sequence, which is located on chromosome 2 of most isolates, was deleted in the cloned parasite line E12 of the FCQ27/PNG isolate. Here we show that erythrocytes infected with the SD17-containing cloned line D10 have typical knob structures on their surfaces, whereas those infected with the line E12 lack knobs. An expression clone was constructed from SD17 and used to affinity purify antibodies from the sera of individuals living in areas of Papua New Guinea where malaria is endemic. The antibodies reacted in immunoblotting experiments with a single polypeptide that varied in Mr from 85,000 to 105,000 among different isolates. The antigen was not expressed in the knobless clone E12. Postembedding immunoelectron microscopy showed localization of the antigen over the knobs of FC27 and two other isolates, largely on the cytoplasmic side. We conclude that the parasite antigen corresponding to clone SD17 is a knob protein. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3542549 ID - 267 ER - TY - JOUR AU - Coppel, R. L. AU - Bianco, A. E. AU - Culvenor, J. G. AU - Crewther, P. E. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1987 TI - A cDNA clone expressing a rhoptry protein of Plasmodium falciparum SP - 73-81 N1 - Aug JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 25 IS - 1 SN - 0166-6851 (Print) N1 - A cDNA clone expressing a rhoptry protein of Plasmodium falciparum N1 - 2823136 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence Chromatography, Affinity Cloning, Molecular DNA/*genetics DNA Restriction Enzymes Deoxyribonuclease EcoRI Deoxyribonuclease HindIII Fluorescent Antibody Technique Genes Humans Immunoassay Microscopy, Electron Molecular Sequence Data Nucleic Acid Hybridization Plasmodium falciparum/*genetics/immunology/ultrastructure N2 - Antibodies from immune humans were used to select a cDNA clone expressing an asexual blood stage antigen of Plasmodium falciparum. The expressed fused polypeptide was used as an affinity reagent to purify human antibodies specific for the corresponding parasite antigen. Western blotting and immunoelectronmicroscopy demonstrated that the antigen was a 105 kDa protein located in the rhoptries of merozoites. The cDNA encodes the carboxy terminus of the rhoptry antigen, a sequence rich both in charged and hydroxy amino acids. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2823136 ID - 263 ER - TY - JOUR AU - Brown, H. AU - Kemp, D. J. AU - Barzaga, N. AU - Brown, G. V. AU - Anders, R. F. AU - Coppel, R. L. PY - 1987 TI - Sequence variation in S-antigen genes of Plasmodium falciparum SP - 365-76 N1 - Dec JF - Mol Biol Med JO - Molecular biology & medicine VL - 4 IS - 6 SN - 0735-1313 (Print) N1 - Sequence variation in S-antigen genes of Plasmodium falciparum N1 - 3325726 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence DNA, Recombinant Genes Molecular Sequence Data Plasmodium falciparum/*genetics Repetitive Sequences, Nucleic Acid Sequence Homology, Nucleic Acid Variation (Genetics) N2 - S-antigens are soluble heat-stable antigens released into the circulation at the time of schizogony of Plasmodium falciparum. Many serologically distinct S-antigens exist and we have shown that this diversity results from repetitive sequences that vary in repeat number, length, sequence and/or reading frame among different S-antigens. We present here the complete sequence of the S-antigen of a Vietnamese isolate V1. The major repeat of 33 base-pairs can be considered to be derived by a deletion event from a 45 base-pair sequence that is located at the 3' repeat boundary and is related in sequence to all S-antigen repeats known so far. We also show that the non-repetitive coding region of the S-antigen gene of V1 is identical to that of K1 and only two amino acids different to that of NF7. In contrast, the sequences are considerably different to those of the FC27 and Wellcome isolates. We conclude that S-antigen genes are highly polymorphic in the repetitive regions but show more restricted diversity in non-repetitive regions. AD - Walter and Eliza Hall Institute of Medical Research, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3325726 ID - 260 ER - TY - JOUR AU - Bianco, A. E. AU - Culvenor, J. G. AU - Coppel, R. L. AU - Crewther, P. E. AU - McIntyre, P. AU - Favaloro, J. M. AU - Brown, G. V. AU - Kemp, D. J. AU - Anders, R. F. PY - 1987 TI - Putative glycophorin-binding protein is secreted from schizonts of Plasmodium falciparum SP - 91-102 N1 - Feb JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 23 IS - 1 SN - 0166-6851 (Print) N1 - Putative glycophorin-binding protein is secreted from schizonts of Plasmodium falciparum N1 - 3553939 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies/immunology Antigens, Protozoan/*analysis/genetics Base Sequence Cloning, Molecular DNA/analysis Electrophoresis, Agar Gel Erythrocytes/parasitology/ultrastructure Fluorescent Antibody Technique Humans Immunologic Techniques Microscopy, Electron Nucleic Acid Hybridization Plasmodium falciparum/genetics/*immunology/ultrastructure N2 - A cDNA clone expressing an antigen of Plasmodium falciparum, selected by screening an expression library cloned in Escherichia coli, encodes a portion of the protein identified as a glycophorin-binding protein [Kochan et al. (1986) Cell 44, 689-696]. Human antibodies affinity-purified on extracts from this clone were used to characterize the antigen by immunoblotting. This protein was present in all isolates tested, restricted to mature trophozoites and schizonts. It was abundant in culture supernatants at the time of merozoite release but present in minor amounts if at all in merozoites. The pattern of antigen distribution over schizont-infected cells observed by immunoelectron microscopy differed from that of the precursor of the major merozoite surface antigens in that most of the antigen appeared to be located over the erythrocyte cytoplasm without any obvious association with organelles. It thus appears unlikely that this antigen is present on the merozoite surface prior to schizont rupture. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3553939 ID - 266 ER - TY - JOUR AU - Stahl, H. D. AU - Bianco, A. E. AU - Crewther, P. E. AU - Burkot, T. AU - Coppel, R. L. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1986 TI - An asparagine-rich protein from blood stages of Plasmodium falciparum shares determinants with sporozoites SP - 3089-102 N1 - Apr 11 JF - Nucleic Acids Res JO - Nucleic acids research VL - 14 IS - 7 SN - 0305-1048 (Print) N1 - An asparagine-rich protein from blood stages of Plasmodium falciparum shares determinants with sporozoites N1 - 2421257 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*analysis Asparagine/*analysis/immunology Base Sequence Cross Reactions DNA/analysis Epitopes/analysis Fluorescent Antibody Technique Humans Invertebrate Hormones/*analysis/immunology Nucleic Acid Hybridization Plasmodium falciparum/*analysis RNA, Messenger/metabolism Repetitive Sequences, Nucleic Acid N2 - We describe a cDNA clone derived from mRNA of asexual blood-stages of the malaria parasite Plasmodium falciparum. This clone, designated Ag319, expresses a P.falciparum antigen fused to beta-galactosidase in Escherichia coli. Human antibodies from Papua New Guinea were affinity-purified by adsorption to extracts of Ag319 immobilized on CNBr-Sepharose. The antibodies reacted predominantly with P. falciparum polypeptides of Mr 220,000 and 160,000, and a number of ill-defined lower molecular weight species. Antibodies reacted in indirect immunofluorescence with all asexual blood-stages although the antigen appeared to be most abundance in the schizont. Surprizingly the antibodies also reacted with sporozoites. The amino acid sequence predicted from the complete nucleotide sequence of this clone is remarkable because 40% of the residues are Asn, and so the antigen has been termed the Asparagine-Rich Protein (ARP). Like other P. falciparum antigens, ARP contains tandemly repetitive sequences, based on the tetrapeptide Asn-Asn-Asn-Met and we have confirmed that these represent natural epitopes by reaction of the corresponding synthetic peptides with human antibodies. Surprisingly, ARP is also rich in Asn outside the tandem repeats. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2421257 ID - 277 ER - TY - JOUR AU - Stahl, H. D. AU - Bianco, A. E. AU - Crewther, P. E. AU - Anders, R. F. AU - Kyne, A. P. AU - Coppel, R. L. AU - Mitchell, G. F. AU - Kemp, D. J. AU - Brown, G. V. PY - 1986 TI - Sorting large numbers of clones expressing Plasmodium falciparum antigens in Escherichia coli by differential antibody screening SP - 351-68 N1 - Aug JF - Mol Biol Med JO - Molecular biology & medicine VL - 3 IS - 4 SN - 0735-1313 (Print) N1 - Sorting large numbers of clones expressing Plasmodium falciparum antigens in Escherichia coli by differential antibody screening N1 - 3534513 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antibodies Antigen-Antibody Complex/analysis Antigens, Protozoan/analysis/*genetics Base Sequence Escherichia coli/*genetics Fluorescent Antibody Technique Humans Malaria/blood Nucleic Acid Hybridization Plasmodium falciparum/classification/*genetics N2 - We describe an approach to classifying a large number of clones expressing Plasmodium falciparum antigens in Escherichia coli by virtue of their differing reactivities with 100 human anti-malarial sera. Individual sera exhibited marked differences in the patterns of reactivity with these clones. These patterns led to the identification of sets of clones, here termed "serological families", which were shown to encode distinct P. falciparum antigens. A serological family was found to be composed of non-identical clones derived from portions of the same antigen. Using this approach six new P. falciparum antigens were identified. One of these is described in detail and is a 102 X 10(3) Mr antigen, predominantly of schizonts. Sequencing studies on four cDNA clones encoding parts of this antigen revealed blocks of hydrophilic dipeptide and tripeptide repeats and so the antigen has been termed the acidic basic repeat antigen (ABRA). UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3534513 ID - 276 ER - TY - JOUR AU - Kemp, D. J. AU - Coppel, R. L. AU - Stahl, H. D. AU - Bianco, A. E. AU - Corcoran, L. M. AU - McIntyre, P. AU - Langford, C. J. AU - Favaloro, J. M. AU - Crewther, P. E. AU - Brown, G. V. AU - et al. PY - 1986 TI - The Wellcome Trust lecture. Genes for antigens of Plasmodium falciparum SP - S83-108 JF - Parasitology JO - Parasitology VL - 92 Suppl SN - 0031-1820 (Print) N1 - The Wellcome Trust lecture. Genes for antigens of Plasmodium falciparum N1 - 2423947 N1 - Journal Article Research Support, Non-U.S. Gov't Review England N1 - eng KW - Amino Acid Sequence Animals Antibody Formation Antigens, Protozoan/*genetics/immunology Chromosome Mapping Cloning, Molecular Cross Reactions DNA/genetics Epitopes/immunology Erythrocyte Membrane/immunology Female Humans Malaria/*immunology/prevention & control Male Nucleic Acid Hybridization Plasmodium falciparum/*genetics/immunology RNA, Messenger/genetics Repetitive Sequences, Nucleic Acid Sequence Homology, Nucleic Acid Vaccines/immunology UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2423947 ID - 282 ER - TY - JOUR AU - Favaloro, J. M. AU - Coppel, R. L. AU - Corcoran, L. M. AU - Foote, S. J. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1986 TI - Structure of the RESA gene of Plasmodium falciparum SP - 8265-77 N1 - Nov 11 JF - Nucleic Acids Res JO - Nucleic acids research VL - 14 IS - 21 SN - 0305-1048 (Print) N1 - Structure of the RESA gene of Plasmodium falciparum N1 - 3537955 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Antigens, Surface/*genetics Base Sequence Cloning, Molecular *Genes Plasmodium falciparum/*genetics/immunology Polymorphism, Genetic *Protozoan Proteins N2 - We have determined the nucleotide sequence of the gene encoding the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum, an antigen that has been shown to confer protective immunity on monkeys. The sequence has enabled us to predict the structure of the RESA gene and the amino acid sequence of its protein product. The gene consists of two exons with a short intron located near the 5' end of the coding region. A hydrophobic amino acid segment predicted for the 3' end of exon 1 is consistent with the possibility that exon 1 encodes trafficking signal sequences. We show that restriction fragment length polymorphisms can be used to define two different alleles of RESA, represented by isolates FC27 and NF7, and compare the FC27 sequence with that of a long cDNA clone from NF7 described previously. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3537955 ID - 271 ER - TY - JOUR AU - Crewther, P. E. AU - Bianco, A. E. AU - Brown, G. V. AU - Coppel, R. L. AU - Stahl, H. D. AU - Kemp, D. J. AU - Anders, R. F. PY - 1986 TI - Affinity purification of human antibodies directed against cloned antigens of Plasmodium falciparum SP - 257-64 N1 - Feb 12 JF - J Immunol Methods JO - Journal of immunological methods VL - 86 IS - 2 SN - 0022-1759 (Print) N1 - Affinity purification of human antibodies directed against cloned antigens of Plasmodium falciparum N1 - 3511156 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Animals Antibodies/*isolation & purification Antibody Specificity Antigens, Protozoan/*immunology Chromatography, Affinity Erythrocytes/immunology Humans Malaria/*immunology Molecular Weight Plasmodium falciparum/growth & development/*immunology N2 - A technique has been developed for the affinity purification of antibodies recognizing cloned antigens of the malaria parasite Plasmodium falciparum expressed in bacteria. Adsorbents prepared by coupling bacterial lysates to Sepharose were used to isolate monospecific antibodies from human immune sera. Production of an abundant stable fused polypeptide by the bacteria was not a prerequisite for the success of this approach. Also the procedure permits the characterization of antigens which elicit the production of very low levels of antibodies. Affinity-purified human antibodies were used to characterized the corresponding P. falciparum antigens by immunoblotting and a number of antigens identified in this way illustrate some commonly observed features of P. falciparum antigens. Several of these antibody preparations recognized multiple bands in the electrophoretic patterns. Studies on a number of isolates of P. falciparum indicate that many antigens exhibit size polymorphisms. Production of some antigens was shown to be restricted to particular stages of the asexual blood cycle of the parasite while others appear to be specifically processed during the life cycle. Affinity-purified antibodies have also been used to locate antigens within the infected erythrocyte and to delineate subsets of antibodies recognizing different epitopes of a single antigen. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3511156 ID - 280 ER - TY - JOUR AU - Coppel, R. L. AU - Culvenor, J. G. AU - Bianco, A. E. AU - Crewther, P. E. AU - Stahl, H. D. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1986 TI - Variable antigen associated with the surface of erythrocytes infected with mature stages of Plasmodium falciparum SP - 265-77 N1 - Sep JF - Mol Biochem Parasitol JO - Molecular and biochemical parasitology VL - 20 IS - 3 SN - 0166-6851 (Print) N1 - Variable antigen associated with the surface of erythrocytes infected with mature stages of Plasmodium falciparum N1 - 3531849 N1 - Journal Article Research Support, Non-U.S. Gov't Netherlands N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*analysis/genetics Antigens, Surface/*analysis/genetics Base Sequence DNA/analysis Electrophoresis, Polyacrylamide Gel Erythrocyte Membrane/immunology/ultrastructure Erythrocytes/immunology/*parasitology/ultrastructure Humans Immune Sera/immunology Immunologic Techniques Microscopy, Electron Nucleic Acid Hybridization Plasmodium falciparum/genetics/*immunology/ultrastructure Repetitive Sequences, Nucleic Acid N2 - Immune human sera were used to select a cDNA clone expressing an asexual blood-stage antigen of Plasmodium falciparum. Antibodies affinity-purified on extracts from this clone were used to characterize the antigen by immunoblotting and immunofluorescence. The antigen is present in mature-stage parasites as a high molecular weight protein of about 250 kDa and is apparently processed to smaller fragments in the merozoite. It varies in molecular weight and antibody reactivity in different isolates, and has been localized at the erythrocyte membrane by immunoelectronmicroscopy. Part of the protein is composed of exactly repeated hexapeptide units that constitute the strain-specific determinant. This molecule has similar characteristics to the strain-specific molecule believed to be responsible for cytoadherence. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3531849 ID - 275 ER - TY - JOUR AU - Coppel, R. L. AU - Crewther, P. E. AU - Favaloro, J. M. AU - Stahl, H. D. PY - 1986 TI - Cloning of asexual blood-stage antigens of Plasmodium falciparum SP - 75-80 N1 - Mar JF - P N G Med J JO - Papua and New Guinea medical journal VL - 29 IS - 1 SN - 0031-1480 (Print) N1 - Cloning of asexual blood-stage antigens of Plasmodium falciparum N1 - 3463017 N1 - Journal Article Research Support, Non-U.S. Gov't Papua new guinea N1 - eng KW - Antigens, Protozoan/*immunology Antigens, Surface/immunology *Cloning, Molecular Erythrocyte Membrane/immunology Humans Malaria/*immunology Molecular Weight UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3463017 ID - 279 ER - TY - JOUR AU - Coppel, R. L. PY - 1986 TI - Prospects for a malaria vaccine SP - 292-5 N1 - Oct JF - Microbiol Sci JO - Microbiological sciences VL - 3 IS - 10 SN - 0265-1351 (Print) N1 - Prospects for a malaria vaccine N1 - 3153563 N1 - Journal Article Research Support, Non-U.S. Gov't Review United states N1 - eng KW - Animals Humans Malaria/*prevention & control Plasmodium falciparum/*immunology *Protozoan Vaccines N2 - Malaria infection is a worsening problem throughout the developing world where conventional methods of control and treatment are becoming ineffective. Recent discoveries using the tools of the new biology, monoclonal antibodies and gene cloning, have brought malaria vaccines to the brink of reality. AD - Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3153563 ID - 273 ER - TY - JOUR AU - Collins, W. E. AU - Anders, R. F. AU - Pappaioanou, M. AU - Campbell, G. H. AU - Brown, G. V. AU - Kemp, D. J. AU - Coppel, R. L. AU - Skinner, J. C. AU - Andrysiak, P. M. AU - Favaloro, J. M. AU - et al. PY - 1986 TI - Immunization of Aotus monkeys with recombinant proteins of an erythrocyte surface antigen of Plasmodium falciparum SP - 259-62 N1 - Sep 18-24 JF - Nature JO - Nature VL - 323 IS - 6085 SN - 0028-0836 (Print) N1 - Immunization of Aotus monkeys with recombinant proteins of an erythrocyte surface antigen of Plasmodium falciparum N1 - 2429187 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antibody Specificity Antigens, Protozoan/genetics/*immunology Antigens, Surface/genetics/immunology Aotus trivirgatus Epitopes Erythrocyte Membrane/*immunology Immunization Plasmodium falciparum/*immunology Recombinant Fusion Proteins/immunology Vaccines, Synthetic N2 - Recent studies have identified and characterized a ring-infected erythrocyte surface antigen (RESA) of the human malaria parasite Plasmodium falciparum with a relative molecular mass (Mr) of approximately 155,000 (refs 1-7). RESA is localized in the micronemes of merozoites and also the membrane of red cells infected with ring-stage parasites. It is thought to be released through the apical pore from the rhoptry at the time of merozoite invasion. Because antibodies directed against this antigen strongly inhibit parasite growth in vitro, RESA may be useful in developing a vaccine against this parasite Here we describe an immunization trial using Aotus monkeys and Escherichia coli-derived fused polypeptides corresponding to various regions of the RESA molecule. Some monkeys in all test groups, but not in the control group, were protected against overwhelming infection. Strikingly, protection correlated with antibody responses to either of two different repetitive sequences in RESA. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2429187 ID - 274 ER - TY - JOUR AU - Bianco, A. E. AU - Favaloro, J. M. AU - Burkot, T. R. AU - Culvenor, J. G. AU - Crewther, P. E. AU - Brown, G. V. AU - Anders, R. F. AU - Coppel, R. L. AU - Kemp, D. J. PY - 1986 TI - A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster SP - 8713-7 N1 - Nov JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 83 IS - 22 SN - 0027-8424 (Print) N1 - A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster N1 - 3095842 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*analysis/genetics/immunology DNA/analysis Drosophila melanogaster/*analysis Heat-Shock Proteins/*analysis Plasmodium falciparum/*analysis/immunology N2 - We describe an antigen of Plasmodium falciparum, defined by a cDNA clone designated Ag63. The antigen is an abundant, soluble cytoplasmic polypeptide of Mr 75,000 present in all stages of asexual development in the blood and in gametocytes, but not in sporozoites. The sequence of the cDNA clone revealed that, like many other antigens of P. falciparum, it contains tandemly repeated amino acid sequences, in this case Gly-Gly-Met-Pro. However, the rest of the sequence is 70% homologous at the amino acid level to the heat shock protein hsp70 of Drosophila melanogaster. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3095842 ID - 272 ER - TY - JOUR AU - Anders, R. F. AU - Shi, P. T. AU - Scanlon, D. B. AU - Leach, S. J. AU - Coppel, R. L. AU - Brown, G. V. AU - Stahl, H. D. AU - Kemp, D. J. PY - 1986 TI - Antigenic repeat structures in proteins of Plasmodium falciparum SP - 164-83 JF - Ciba Found Symp JO - Ciba Foundation symposium VL - 119 SN - 0300-5208 (Print) N1 - Antigenic repeat structures in proteins of Plasmodium falciparum N1 - 2426051 N1 - Journal Article Research Support, Non-U.S. Gov't Review Netherlands N1 - eng KW - Amino Acid Sequence Animals Antibodies, Monoclonal/immunology Antigens, Protozoan/*immunology Antigens, Surface/immunology Cross Reactions Epitopes/immunology Erythrocytes/immunology Malaria/immunology Peptides/immunology Plasmodium falciparum/*immunology *Protozoan Proteins N2 - The majority of malaria antigens that have been cloned contain short sequence repeats which encode antigenic epitopes that are naturally immunogenic. Synthetic peptides have been used to show that natural antibody responses to a strain-specific Plasmodium falciparum S antigen are largely directed against epitopes encoded in an 11-amino acid sequence that is repeated approximately 100 times in the molecule. A 16-amino acid peptide conjugated to bovine serum albumin induced antibodies specific for the S antigen of the homologous isolate. Synthetic peptides have also been used to confirm the natural immunogenicity of epitopes encoded within two blocks of related repeats in the Ring-infected Erythrocyte Surface Antigen (RESA). A 16-amino acid peptide, comprising four repeats of the tetrameric sequence EENV, induced antibodies reactive with the native molecule. Detailed analyses of these anti-peptide antisera indicate that short sequence repeats express more than one epitope, some of which may cross-react with other repeat structures. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2426051 ID - 281 ER - TY - JOUR AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. AU - Kemp, D. J. PY - 1986 TI - Repeat structures in malaria antigens SP - 87-93 N1 - Mar JF - P N G Med J JO - Papua and New Guinea medical journal VL - 29 IS - 1 SN - 0031-1480 (Print) N1 - Repeat structures in malaria antigens N1 - 3463018 N1 - Journal Article Research Support, Non-U.S. Gov't Papua new guinea N1 - eng KW - Amino Acid Sequence Antigens, Protozoan/*analysis Humans Malaria/*immunology Peptides/*analysis UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3463018 ID - 278 ER - TY - JOUR AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. AU - Kemp, D. J. PY - 1986 TI - Induction of protective immunity to malaria SP - 245-53 N1 - Dec JF - Lepr Rev JO - Leprosy review VL - 57 Suppl 2 SN - 0305-7518 (Print) N1 - Induction of protective immunity to malaria N1 - 3553798 N1 - Journal Article England N1 - eng KW - Animals Antigens, Protozoan/genetics/*immunology Antigens, Surface/genetics/immunology Aotus trivirgatus Cloning, Molecular Erythrocytes/parasitology Malaria/immunology/*prevention & control Plasmodium falciparum/genetics/immunology *Protozoan Proteins Vaccines, Synthetic/genetics/immunology UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3553798 ID - 270 ER - TY - JOUR AU - Stahl, H. D. AU - Kemp, D. J. AU - Crewther, P. E. AU - Scanlon, D. B. AU - Woodrow, G. AU - Brown, G. V. AU - Bianco, A. E. AU - Anders, R. F. AU - Coppel, R. L. PY - 1985 TI - Sequence of a cDNA encoding a small polymorphic histidine- and alanine-rich protein from Plasmodium falciparum SP - 7837-46 N1 - Nov 11 JF - Nucleic Acids Res JO - Nucleic acids research VL - 13 IS - 21 SN - 0305-1048 (Print) N1 - Sequence of a cDNA encoding a small polymorphic histidine- and alanine-rich protein from Plasmodium falciparum N1 - 2415925 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antibodies Antigens, Protozoan/*genetics/immunology Base Sequence *Cloning, Molecular DNA/*metabolism Epitopes/analysis Escherichia coli/genetics Fluorescent Antibody Technique Humans Plasmodium falciparum/*genetics *Polymorphism, Genetic Protein Biosynthesis Repetitive Sequences, Nucleic Acid N2 - We describe the expression in Escherichia coli, isolation by immunological screening and complete nucleotide sequence of a cDNA clone from the malaria parasite Plasmodium falciparum. The deduced amino acid sequence contains separate blocks of repetitive hexapeptide and pentapeptide sequences and we have confirmed that these represent epitopes by reaction of the corresponding synthetic peptides with human antibodies. As the predicted size is Mr 21,000 and the overall composition is 30% His and 29% Ala, the polypeptide has been termed the small histidine-alanine rich protein (SHARP). This polypeptide is highly polymorphic in different P. falciparum isolates and cross reacts immunologically with a distinct gene product of P. falciparum. Although it is related to the Histidine Rich Protein (HRP) of P. lophurae by virtue of its high His content, it shows no obvious sequence relationship to the HRP outside the repeats. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2415925 ID - 283 ER - TY - JOUR AU - Stahl, H. D. AU - Crewther, P. E. AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. AU - Bianco, A. E. AU - Mitchell, G. F. AU - Kemp, D. J. PY - 1985 TI - Interspersed blocks of repetitive and charged amino acids in a dominant immunogen of Plasmodium falciparum SP - 543-7 N1 - Jan JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 82 IS - 2 SN - 0027-8424 (Print) N1 - Interspersed blocks of repetitive and charged amino acids in a dominant immunogen of Plasmodium falciparum N1 - 3881769 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Amino Acids/*analysis Animals Antibodies/immunology Antigens, Protozoan/analysis/*genetics Base Sequence DNA/analysis Escherichia coli Humans Molecular Weight Plasmodium falciparum/*immunology Repetitive Sequences, Nucleic Acid N2 - We describe an antigen of Plasmodium falciparum that is a dominant immunogen in man. The corresponding cDNA clone, Ag231, expressing this antigen in Escherichia coli reacted in an in situ colony assay with sera from up to approximately equal to 93% of 65 people living in an area in which P. falciparum is endemic. Human antibodies affinity purified on immobilized Ag231 lysates identified the corresponding parasite antigen as a polypeptide of Mr approximately equal to 300,000. It was present in schizonts and also in ring-stage trophozoites, where a speckled immunofluorescence pattern suggested an association with the erythrocyte. Its mRNA was enriched in merozoites relative to other blood stages, a distinctive property shared by a recently described antigen located on the surface of ring-infected erythrocytes, and it is encoded by a single gene having a number of allelic variants. The complete nucleotide sequence of Ag231 revealed a structural unit composed of 13 hexapeptide repeats flanked by a highly charged region containing both acidic and basic amino acids. This structural unit is itself repeated, so that blocks of repeats and charged units are interspersed along the molecule. The sequences within the repeats vary much more extensively than those in the charged units. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3881769 ID - 289 ER - TY - JOUR AU - Kemp, D. J. AU - Corcoran, L. M. AU - Coppel, R. L. AU - Stahl, H. D. AU - Bianco, A. E. AU - Brown, G. V. AU - Anders, R. F. PY - 1985 TI - Size variation in chromosomes from independent cultured isolates of Plasmodium falciparum SP - 347-50 N1 - May 23-29 JF - Nature JO - Nature VL - 315 IS - 6017 SN - 0028-0836 (Print) N1 - Size variation in chromosomes from independent cultured isolates of Plasmodium falciparum N1 - 3889657 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antigens, Protozoan/genetics Chromosomes/*ultrastructure DNA/isolation & purification Electrophoresis, Agar Gel/methods Molecular Weight Plasmodium falciparum/*genetics Repetitive Sequences, Nucleic Acid Species Specificity N2 - The complexity of the life cycle of the protozoan malaria parasite Plasmodium falciparum has hindered genetic analysis; even the number of chromosomes in P. falciparum is uncertain. The blood stages of rodent malaria parasites are haploid and hybridization with cloned complementary DNAs similarly suggests a haploid genome in P. falciparum blood stages (ref. 4 and our unpublished results). A novel approach to karyoptic and linkage analysis in P. falciparum has been provided recently by the technique of pulsed-field gradient (PFG) gel electrophoresis, which allows the fractionation of DNA molecules of 30-3,000 kilobases (kb), a range including the sizes of intact chromosomal DNA molecules from eukaryotes such as yeast and trypanosomatids. We describe here the fractionation by PFG electrophoresis of chromosomal DNA molecules from P. falciparum into at least seven discrete species which vary in size by up to 20% between different isolates. Several genes for P. faciparum antigens which contain repetitive sequences are located on different chromosomes. Surprisingly, two of the chromosomes seem to contain the same sequences. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3889657 ID - 287 ER - TY - JOUR AU - Cowman, A. F. AU - Saint, R. B. AU - Coppel, R. L. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1985 TI - Conserved sequences flank variable tandem repeats in two S-antigen genes of Plasmodium falciparum SP - 775-83 N1 - Apr JF - Cell JO - Cell VL - 40 IS - 4 SN - 0092-8674 (Print) N1 - Conserved sequences flank variable tandem repeats in two S-antigen genes of Plasmodium falciparum N1 - 3886159 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Composition Base Sequence Codon *Dna DNA, Recombinant Nucleic Acid Hybridization Peptide Fragments Peptides Plasmodium falciparum/*genetics/immunology Protein Sorting Signals Repetitive Sequences, Nucleic Acid N2 - We describe the isolation of two chromosomal DNA fragments from Plasmodium falciparum. These fragments encode the antigenically distinct S antigens of two different P. falciparum isolates, namely FC27 from Papua New Guinea and NF7 from Ghana. The complete nucleotide sequences of both fragments are presented. The fragments are homologous over most of their lengths, including the entire regions flanking the protein coding sequences. Whereas the N- and C-terminal portions of sequences encoding the S antigens are homologous, major portions of the coding sequences are not. The nonhomologous regions are comprised of tandemly repeated sequences, of 33 bp in FC27 and predominantly of 24 bp in NF7. The 33 bp tandem repeats encoded by the FC27 S-antigen gene could not be detected in the NF7 genome. Conversely, the 24 bp tandem repeats encoded by the NF7 S-antigen gene could not be detected in the FC27 genome. The pattern of sequence variation within the repeats of both genes suggests a mechanism for the generation of S-antigen diversity. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3886159 ID - 288 ER - TY - JOUR AU - Coppel, R. L. AU - Saint, R. B. AU - Stahl, H. D. AU - Langford, C. J. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1985 TI - Plasmodium falciparum: differentiation of isolates with DNA hybridization using antigen gene probes SP - 82-9 N1 - Aug JF - Exp Parasitol JO - Experimental parasitology VL - 60 IS - 1 SN - 0014-4894 (Print) N1 - Plasmodium falciparum: differentiation of isolates with DNA hybridization using antigen gene probes N1 - 2990991 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens, Protozoan/*genetics Autoradiography Collodion Dna DNA Restriction Enzymes Deoxyribonuclease EcoRI Deoxyribonuclease HindIII Electrophoresis, Agar Gel Nucleic Acid Hybridization Plasmodium falciparum/*classification/genetics/immunology Polymorphism, Genetic N2 - Chromosomal DNA was prepared from seven Plasmodium falciparum isolates that had been cultured in vitro and from a cloned P. falciparum line. The DNA was cleaved with restriction endonucleases, fractionated by agarose gel electrophoresis, blotted to nitrocellulose, and hybridized with a series of radioactively labeled DNA probes. The probes had been derived from cDNA clones encoding portions of P. falciparum antigens. Simple, reproducible band patterns that differed for many of the isolates were obtained. Parasite isolates collected from different continents could be readily distinguished, as could some but not all isolates collected from one restricted region of Papua New Guinea. Application of this technique for the identification and differentiation of parasite strains was explored. The patterns of hybridization observed were consistent with the proposition that blood stages of P. falciparum have a haploid genome. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2990991 ID - 285 ER - TY - JOUR AU - Coppel, R. L. AU - Favaloro, J. M. AU - Crewther, P. E. AU - Burkot, T. R. AU - Bianco, A. E. AU - Stahl, H. D. AU - Kemp, D. J. AU - Anders, R. F. AU - Brown, G. V. PY - 1985 TI - A blood stage antigen of Plasmodium falciparum shares determinants with the sporozoite coat protein SP - 5121-5 N1 - Aug JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 82 IS - 15 SN - 0027-8424 (Print) N1 - A blood stage antigen of Plasmodium falciparum shares determinants with the sporozoite coat protein N1 - 2410913 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Amino Acid Sequence Animals Antigens, Protozoan/genetics/*immunology Base Sequence Epitopes Humans Membrane Proteins/genetics/immunology Plasmodium falciparum/genetics/*immunology N2 - A cDNA clone expressing a Plasmodium falciparum blood-stage antigen in Escherichia coli was identified by colony immunoassay using immune human sera. Antibodies affinity-purified on extracts of this clone reacted with both asexual blood stages and sporozoites of P. falciparum, recognizing a Mr23,000 protein in the blood stages. The nucleotide sequence of the cDNA revealed a signal peptide and an internal hydrophobic sequence typical of transmembrane anchor sequences. Located 3' to the putative anchor are two tetramers, Asn-Ala-Asn-Pro and Asn-Ala-Asp-Pro, which are closely related to the repeats of the circumsporozoite protein of P. falciparum. The blood stage protein is conserved amongst several isolates of P. falciparum, and antibodies against it are common in the sera of individuals living in the area where the parasite is endemic. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2410913 ID - 286 ER - TY - JOUR AU - Brown, G. V. AU - Culvenor, J. G. AU - Crewther, P. E. AU - Bianco, A. E. AU - Coppel, R. L. AU - Saint, R. B. AU - Stahl, H. D. AU - Kemp, D. J. AU - Anders, R. F. PY - 1985 TI - Localization of the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum in merozoites and ring-infected erythrocytes SP - 774-9 N1 - Aug 1 JF - J Exp Med JO - The Journal of experimental medicine VL - 162 IS - 2 SN - 0022-1007 (Print) N1 - Localization of the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum in merozoites and ring-infected erythrocytes N1 - 3894564 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens, Protozoan/*isolation & purification Antigens, Surface/isolation & purification Erythrocytes/immunology/parasitology/ultrastructure Humans Malaria/blood/*immunology/parasitology Microscopy, Electron Plasmodium falciparum/growth & development/*immunology/ultrastructure N2 - Immunoelectron microscopy with protein A gold has been used to determine the subcellular location of the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum. RESA was associated with dense vesicles presumed to be micronemes within merozoites. RESA was not detected on the surface of merozoites but was located at the membrane of erythrocytes infected with ring-stage parasites. RESA within merozoites was largely soluble in the nonionic detergent Triton X-100, but was insoluble in this detergent when associated with the erythrocyte membrane. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3894564 ID - 284 ER - TY - JOUR AU - Anders, R. F. AU - Brown, G. V. AU - Coppel, R. L. AU - Stahl, H. D. AU - Bianco, A. E. AU - Favaloro, J. M. AU - Crewther, P. E. AU - Culvenor, J. G. AU - Kemp, D. J. PY - 1985 TI - Potential vaccine antigens of the asexual blood-stages of Plasmodium falciparum SP - 81-9 JF - Dev Biol Stand JO - Developments in biological standardization VL - 62 SN - 0301-5149 (Print) N1 - Potential vaccine antigens of the asexual blood-stages of Plasmodium falciparum N1 - 2422079 N1 - Journal Article Research Support, Non-U.S. Gov't Switzerland N1 - eng KW - Animals Antibodies/immunology Antigens, Protozoan/genetics/*immunology Antigens, Surface/immunology Cloning, Molecular DNA/genetics DNA, Recombinant Epitopes/immunology Erythrocytes/parasitology Escherichia coli/genetics/immunology Humans Malaria/*immunology/parasitology/prevention & control Plasmodium falciparum/genetics/*immunology Repetitive Sequences, Nucleic Acid Vaccines/*immunology N2 - We have constructed a cDNA clone library that contains many natural immunogens of the asexual blood-stages of Plasmodium falciparum. The corresponding parasite antigens have been identified with antisera raised against the antigens expressed in Escherichia coli or with monospecific human antibodies purified on adsorbents prepared from various clones. Sequencing studies on the clones have revealed that many malaria antigens contain extensive sequence repeats. These repeats encode antigenic epitopes that in several proteins have been shown to be immunodominant. One candidate vaccine molecule, the Ring-infected Erythrocyte Surface Antigen (RESA), which is transferred from inside merozoites to the erythrocyte surface at about the time of merozoite invasion, contains two blocks of antigenically cross-reactive repeats. The structures of other antigens containing extensive repeats are described and their possible significance to the host-parasite relationship is discussed. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2422079 ID - 290 ER - TY - JOUR AU - Stahl, H. D. AU - Coppel, R. L. AU - Brown, G. V. AU - Saint, R. AU - Lingelbach, K. AU - Cowman, A. F. AU - Anders, R. F. AU - Kemp, D. J. PY - 1984 TI - Differential antibody screening of cloned Plasmodium falciparum sequences expressed in Escherichia coli: procedure for isolation of defined antigens and analysis of human antisera SP - 2456-60 N1 - Apr JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 81 IS - 8 SN - 0027-8424 (Print) N1 - Differential antibody screening of cloned Plasmodium falciparum sequences expressed in Escherichia coli: procedure for isolation of defined antigens and analysis of human antisera N1 - 6371814 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens/*genetics/isolation & purification Base Sequence *Cloning, Molecular DNA/*metabolism Escherichia coli/*genetics Humans *Immune Sera Immunoassay Plasmodium falciparum/*genetics/immunology N2 - We describe a procedure that uses polyspecific human sera for screening Escherichia coli colonies expressing cloned Plasmodium falciparum cDNA sequences in order to detect colonies that react differentially with different sera. This procedure can be used for two distinct purposes. First, it enables the isolation of clones encoding specified antigenic sequences present in the complex mixture, without purification of either antigens or antibodies by conventional procedures. This requires that the antigen can be expressed in E. coli and that antisera are available that differ substantially in their reactivities to the component of interest. To develop the procedure, we used two polyspecific sera that shared many anti-P. falciparum specificities but differed in that only one was reactive to the isolate-specific S antigen of P. falciparum strain FCQ27/PNG (called FC27). Differential screening with the two sera identified 30 cDNA clones, and colony hybridization confirmed that 25 of these express S-antigen sequences. Second, the procedure identifies defined antibody specificities within polyspecific human sera by virtue of their ability to react with any given cDNA clone. The procedure has been used here to identify antibody specificities that increase dramatically in titer between the acute and convalescent phases of malaria in certain individuals and, hence, to isolate clones encoding the corresponding antigens. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6371814 ID - 295 ER - TY - JOUR AU - Cowman, A. F. AU - Coppel, R. L. AU - Saint, R. B. AU - Favaloro, J. AU - Crewther, P. E. AU - Stahl, H. D. AU - Bianco, A. E. AU - Brown, G. V. AU - Anders, R. F. AU - Kemp, D. J. PY - 1984 TI - The ring-infected erythrocyte surface antigen (RESA) polypeptide of Plasmodium falciparum contains two separate blocks of tandem repeats encoding antigenic epitopes that are naturally immunogenic in man SP - 207-21 N1 - Jun JF - Mol Biol Med JO - Molecular biology & medicine VL - 2 IS - 3 SN - 0735-1313 (Print) N1 - The ring-infected erythrocyte surface antigen (RESA) polypeptide of Plasmodium falciparum contains two separate blocks of tandem repeats encoding antigenic epitopes that are naturally immunogenic in man N1 - 6085696 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Africa Amino Acid Sequence Animals Antigens, Protozoan/*genetics Base Sequence Cloning, Molecular DNA/genetics Epitopes Genes Ghana New Guinea Plasmodium falciparum/genetics/*immunology Repetitive Sequences, Nucleic Acid N2 - We showed previously that the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum contains a repetitive amino acid sequence. We have investigated here the sequence and antigenic relationships between RESA from FC27, a Papua New Guinea isolate, and from NF7, a Ghanaian isolate. The complete nucleotide sequences of eight different cDNA clones demonstrate that RESA from the two strains are closely homologous over the region that can be compared. A series of related eight, four and three amino acid repeats is located at the 3' end, forming the C terminus of RESA in FC27. RESA contains a second block of repeats based on an 11 amino acid sequence and separated from the 3' block by 381 amino acids in NF7. Antibodies from Papua New Guineans react with RESA from the African isolate, and vice-versa. Antigenic determinants that are naturally immunogenic in man are present in the 5' repeats of RESA, as well as in the 3' repeats, and antibodies that cross-react with both blocks of repeats were detected by reacting affinity-purified human antibodies with cloned subfragments of the cDNA clones, expressed in Escherichia coli. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6085696 ID - 294 ER - TY - JOUR AU - Coppel, R. L. AU - Cowman, A. F. AU - Anders, R. F. AU - Bianco, A. E. AU - Saint, R. B. AU - Lingelbach, K. R. AU - Kemp, D. J. AU - Brown, G. V. PY - 1984 TI - Immune sera recognize on erythrocytes Plasmodium falciparum antigen composed of repeated amino acid sequences SP - 789-92 N1 - Aug 30-Sep 5 JF - Nature JO - Nature VL - 310 IS - 5980 SN - 0028-0836 (Print) N1 - Immune sera recognize on erythrocytes Plasmodium falciparum antigen composed of repeated amino acid sequences N1 - 6382025 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals Antigen-Antibody Complex Antigens, Surface/*analysis Erythrocyte Membrane/*immunology Fluorescent Antibody Technique Humans Immune Sera Plasmodium falciparum/*immunology N2 - Protective immune responses against the asexual stages of the human malaria parasite, Plasmodium falciparum, are most probably directed against exposed antigenic determinants on the surface of the free merozoite or the infected red blood cell, and therefore antigens in these locations are candidates for testing as components of a defined molecular vaccine. To facilitate the search for such antigens, we recently developed a method for the expression of P. falciparum proteins in Escherichia coli as fused polypeptides. Many clones producing antigens were detected by screening with immune human sera. We show here that antibodies against the fused polypeptide expressed by one such clone react with a P. falciparum protein that is synthesized late in schizogony and is later present on the surface of the ring-infected erythrocyte. The protein is composed of repeating subunits of 8, 4 and 3 amino acids and is present in all isolates of P. falciparum examined. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6382025 ID - 292 ER - TY - JOUR AU - Coppel, R. L. AU - Brown, G. V. AU - Mitchell, G. F. AU - Anders, R. F. AU - Kemp, D. J. PY - 1984 TI - Identification of a cDNA clone encoding a mature blood stage antigen of Plasmodium falciparum by immunization of mice with bacterial lysates SP - 403-7 N1 - Feb JF - Embo J JO - The EMBO journal VL - 3 IS - 2 SN - 0261-4189 (Print) N1 - Identification of a cDNA clone encoding a mature blood stage antigen of Plasmodium falciparum by immunization of mice with bacterial lysates N1 - 6370681 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antigens, Surface/*genetics Cloning, Molecular *DNA, Recombinant Escherichia coli/genetics Immunization Mice Plasmids Plasmodium falciparum/*genetics/growth & development/immunology N2 - A cDNA library was constructed in pBR322 using mRNA from blood stages of a Papua New Guinean isolate of Plasmodium falciparum. Expression of parasite antigens was not directly detectable by conventional immunological assays. To circumvent this, mice were immunized with lysates of cDNA clones, and the antisera raised were assayed for anti-parasite reactivity. One cDNA clone was identified which reliably elicited antibodies to P. falciparum. The mouse antisera were used to characterize the native P. falciparum protein as a 120-kd protein, which is antigenic during natural infection. The protein occurs in late trophozoite and schizont stages and is found in isolates of the parasite from widely separated geographical areas. The genomic context of the antigen gene is conserved in the different isolates. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6370681 ID - 296 ER - TY - JOUR AU - Brown, G. V. AU - Anders, R. F. AU - Coppel, R. L. AU - Saint, R. B. AU - Cowman, A. F. AU - Stahl, H. D. AU - Lingelbach, K. R. AU - Mitchell, G. F. AU - Alpers, M. P. AU - Kemp, D. J. PY - 1984 TI - The expression of Plasmodium falciparum bloodstage antigens in Escherichia coli SP - 179-87 N1 - Nov 13 JF - Philos Trans R Soc Lond B Biol Sci JO - Philosophical transactions of the Royal Society of London VL - 307 IS - 1131 SN - 0962-8436 (Print) N1 - The expression of Plasmodium falciparum bloodstage antigens in Escherichia coli N1 - 6151682 N1 - Comparative Study Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antigens, Protozoan/*genetics Cloning, Molecular DNA/genetics Escherichia coli/genetics Humans Immunization Malaria/blood/prevention & control Plasmodium falciparum/genetics/growth & development/*immunology Proteins/immunology Repetitive Sequences, Nucleic Acid Vaccines/isolation & purification N2 - A library of cDNA clones expressing proteins of the asexual blood stages of a Papua New Guinean isolate of Plasmodium falciparum (isolate FCQ27/PNG (FC27] was constructed in the bacteriophage vector lambda gt11-Amp3. In an in situ colony immunoassay, human serum was used to identify colonies producing natural immunogens. Sera from donors of defined clinical status, or reactive to a defined subset of natural immunogens were used to identify clones of particular interest (for example, clones reacting with convalescent but not with acute serum or clones expressing the isolate specific S-antigen of FC27). Antisera raised by immunizing mice and rabbits with cloned antigens were used to characterize the P. falciparum proteins corresponding to the antigen-positive clones. Nucleotide sequence analysis of an antigen found on the surface of cells infected with ring stage parasites revealed an unusual sequence coding for eight, four and three amino acid repeats rich in acidic amino acids. The discussion centres on the use of cloned antigens as tools for the analysis of the host-protective immune response and selection of candidate vaccine molecules. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6151682 ID - 291 ER - TY - JOUR AU - Anders, R. F. AU - Coppel, R. L. AU - Brown, G. V. AU - Saint, R. B. AU - Cowman, A. F. AU - Lingelbach, K. R. AU - Mitchell, G. F. AU - Kemp, D. J. PY - 1984 TI - Plasmodium falciparum complementary DNA clones expressed in Escherichia coli encode many distinct antigens SP - 177-91 N1 - Jun JF - Mol Biol Med JO - Molecular biology & medicine VL - 2 IS - 3 SN - 0735-1313 (Print) N1 - Plasmodium falciparum complementary DNA clones expressed in Escherichia coli encode many distinct antigens N1 - 6399547 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Animals Antigens, Protozoan/classification/*genetics Base Sequence DNA/genetics Escherichia coli/genetics Gene Expression Regulation Molecular Weight Plasmodium falciparum/*genetics/growth & development/immunology N2 - A library of cDNA clones expressing antigens of the asexual blood-stages of Plasmodium falciparum (isolate FCQ27/PNG) was constructed in the bacteriophage vector gamma gt11-Amp3. Clones expressing P. falciparum antigens (as polypeptides fused to beta-galactosidase) were selected by their reactivity in an in situ colony immunoassay with affinity-purified malaria antibodies. A detailed analysis of 78 antigen-positive clones selected from approximately 10,000 recombinant clones has shown them to correspond to many different parasite antigens. cDNA hybridization studies on this array of 78 antigen-positive clones have so far identified 18 families of sibling clones with 22 clones as yet unassigned, the majority of which may represent additional unique sequences. Only about 20% of the clones synthesized abundant amounts of the malaria antigen/beta-galactosidase fused polypeptide but each multi-member family except one was represented by at least one clone producing a fused polypeptide in abundance. Antisera have been raised against cloned malaria antigens by immunizing mice and rabbits with bacterial lysates and purified fused polypeptides, respectively. These antisera have been used to characterize the antigens in P. falciparum that correspond to the various antigen-positive clones. The variety of distinct antigens recognized by these antisera confirms that the clone library contains coding sequences for many different antigens. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6399547 ID - 293 ER - TY - JOUR AU - Kemp, D. J. AU - Coppel, R. L. AU - Cowman, A. F. AU - Saint, R. B. AU - Brown, G. V. AU - Anders, R. F. PY - 1983 TI - Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: detection with antibodies from immune humans SP - 3787-91 N1 - Jun JF - Proc Natl Acad Sci U S A JO - Proceedings of the National Academy of Sciences of the United States of America VL - 80 IS - 12 SN - 0027-8424 (Print) N1 - Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: detection with antibodies from immune humans N1 - 6304737 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens/*genetics Base Sequence DNA/analysis DNA Restriction Enzymes Escherichia coli/*genetics Humans Immune Sera/isolation & purification Immunity Malaria/*immunology Plasmids Plasmodium falciparum/genetics/*immunology RNA, Messenger/genetics beta-Galactosidase/genetics N2 - Many proteins produced by blood stages of the malaria parasite Plasmodium falciparum are natural immunogens in man. As an approach to determining which of these are relevant to protective immunity we have constructed an expression library of P. falciparum cDNA sequences, cloned in Escherichia coli. The cDNA sequences were inserted into the beta-galactosidase gene of an ampicillin-resistant derivative of the temperature-sensitive lysogenic bacteriophage lambda gt11. About 5% of the resulting clones expressed P. falciparum sequences as polypeptides fused to beta-galactosidase. We have identified many clones that express P. falciparum antigens by immunological screening in situ with antibodies from immune human sera that inhibit P. falciparum growth in vitro. The antigen-positive clones contain P. falciparum cDNA sequences, as determined by hybridization. Some express polypeptides that are larger than beta-galactosidase and react both with antibodies to beta-galactosidase and with antibodies from humans immune to P. falciparum. The cloned P. falciparum antigens should facilitate new approaches to the identification of potential vaccine molecules. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6304737 ID - 298 ER - TY - JOUR AU - Coppel, R. L. AU - Cowman, A. F. AU - Lingelbach, K. R. AU - Brown, G. V. AU - Saint, R. B. AU - Kemp, D. J. AU - Anders, R. F. PY - 1983 TI - Isolate-specific S-antigen of Plasmodium falciparum contains a repeated sequence of eleven amino acids SP - 751-6 N1 - Dec 22-1984 Jan 4 JF - Nature JO - Nature VL - 306 IS - 5945 SN - 0028-0836 (Print) N1 - Isolate-specific S-antigen of Plasmodium falciparum contains a repeated sequence of eleven amino acids N1 - 6361573 N1 - Journal Article Research Support, Non-U.S. Gov't England N1 - eng KW - Amino Acid Sequence Animals *Antigens, Surface/genetics Base Sequence Genes Isoelectric Point Molecular Weight Plasmodium falciparum/genetics/*immunology Species Specificity N2 - Antibodies raised against a Plasmodium falciparum protein expressed in Escherichia coli reacted with a 220,000-molecular weight antigen of mature blood-stage parasites. The protein resembles the sporozoite surface antigen being composed of tandem repeats of 11 amino acids. However, it is isolate-specific and the encoding gene is not detectable in strains that do not express the protein. UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=6361573 ID - 297 ER - TY - JOUR AU - Brown, G. V. AU - Coppel, R. L. AU - Vrbova, H. AU - Grumont, R. J. AU - Anders, R. F. PY - 1982 TI - Plasmodium falciparum: comparative analysis of erythrocyte stage-dependent protein antigens SP - 279-84 N1 - Apr JF - Exp Parasitol JO - Experimental parasitology VL - 53 IS - 2 SN - 0014-4894 (Print) N1 - Plasmodium falciparum: comparative analysis of erythrocyte stage-dependent protein antigens N1 - 7037443 N1 - Journal Article Research Support, Non-U.S. Gov't United states N1 - eng KW - Animals Antigens/*analysis/isolation & purification Electrophoresis, Polyacrylamide Gel Erythrocytes/*parasitology Humans Molecular Weight Plasmodium falciparum/growth & development/*immunology Proteins/*immunology UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7037443 ID - 299 ER - TY - JOUR AU - Coppel, R. L. AU - Mitchell, G. F. PY - 1975 TI - Studies on accessory cells in the adoptive antibody response to sheep erythrocytes in mice SP - 411-23 N1 - Aug JF - Cell Immunol JO - Cellular immunology VL - 18 IS - 2 SN - 0008-8749 (Print) N1 - Studies on accessory cells in the adoptive antibody response to sheep erythrocytes in mice N1 - 1095217 N1 - Journal Article United states N1 - eng KW - Animals *Antibody Formation Antibody-Producing Cells Antigens Antilymphocyte Serum Cell Adhesion Chromium Radioisotopes Erythrocytes/*immunology Female Hemolytic Plaque Technique Lymph Nodes/cytology Lymphocytes/*immunology Mice Mice, Inbred BALB C Mice, Inbred C57BL Mice, Inbred CBA Mice, Inbred DBA Radiation Chimera Sheep/immunology Spleen/cytology Thymus Gland/cytology UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1095217 ID - 300 ER -