The Experts below are selected from a list of 36 Experts worldwide ranked by ideXlab platform
William H. Robinson - One of the best experts on this subject based on the ideXlab platform.
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Autoantigen microarrays for multiplex Characterization of Autoantibody responses
Nature Medicine, 2002Co-Authors: William H. Robinson, Carla Digennaro, Wolfgang Hueber, Brian B. Haab, Makoto Kamachi, Erik J. Dean, Sylvie Fournel, Derek A. Fong, Mark C. Genovese, Henry E. Neuman De VegvarAbstract:We constructed miniaturized autoantigen arrays to perform large-scale multiplex Characterization of Autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect Autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of Autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases.
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protein microarray Characterization of Autoantibody responses in rheumatoid arthritis and systemic lupus erythematosus
Arthritis Research & Therapy, 2002Co-Authors: William H. Robinson, W Huber, W I Van Venrooij, Sylviane Muller, G S Panayi, C Verweij, Josef S Smolen, Lawrence Steinman, Paul J UtzAbstract:In rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and other autoimmune rheumatic diseases, our understanding of the specificity of the Autoantibody response and the role of autoantibodies in pathogenesis are limited. We developed antigen microarray technology to perform multiplex Characterization of Autoantibody responses. Antigen microarrays are produced by attaching thousands of proteins and peptides to addressable locations on the surface of solid supports using a robotic arrayer. Arrays are probed with serum from disease and control patients, followed by anti-human secondary antibodies covalently-conjugated to spectrally-resolvable fluorochromes. We initially developed a 'connective tissue disease' array containing structurally diverse autoantigens including nucleic acids, histones, hnRNPs, snRNPs, collagens, Ro, La, SCL-70, CENP-B, Jo-1, pyruvate dehydrogenase, and post-translationally-modified antigens. Array analysis of serum derived from patients with SLE, rheumatoid arthritis, Sjogren's syndrome, mixed connective tissue diseases, scleroderma, myositidies, and primary biliary cirrhosis identified Autoantibody response patterns characteristic of these diseases (Fig. (Fig.1).1). We are now developing 'synovial proteome' arrays to study the Autoantibody responses in RA, and have produced first-generation arrays containing 450 distinct protein and peptide candidate antigens derived from synovial joints. These include native and citrulline-modified filaggrin peptides, native and deiminated fibrinogen and vimentin, glucose-6-phosphate isomerase, collagen types I, II, III, IV and V, hnRNP-A2/RA33, hnRNP-D, immunoglobulins, HCgp39, BiP, and HSPs 60, 65, 70, and 90. We probe our arrays with sera and synovial fluid from RA and control patients. We are using our arrays to examine serial serum samples from patients with early RA to: (1) identify antigens targeted early in disease, and (2) examine for evidence of B-cell epitope spreading. 'Synovial proteome' arrays represent a powerful tool to study the breadth and specificity of autoreactive B-cell responses, and to identify candidate and define relevant autoantigens in RA. Figure 1
Paul J Utz - One of the best experts on this subject based on the ideXlab platform.
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protein microarray Characterization of Autoantibody responses in rheumatoid arthritis and systemic lupus erythematosus
Arthritis Research & Therapy, 2002Co-Authors: William H. Robinson, W Huber, W I Van Venrooij, Sylviane Muller, G S Panayi, C Verweij, Josef S Smolen, Lawrence Steinman, Paul J UtzAbstract:In rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and other autoimmune rheumatic diseases, our understanding of the specificity of the Autoantibody response and the role of autoantibodies in pathogenesis are limited. We developed antigen microarray technology to perform multiplex Characterization of Autoantibody responses. Antigen microarrays are produced by attaching thousands of proteins and peptides to addressable locations on the surface of solid supports using a robotic arrayer. Arrays are probed with serum from disease and control patients, followed by anti-human secondary antibodies covalently-conjugated to spectrally-resolvable fluorochromes. We initially developed a 'connective tissue disease' array containing structurally diverse autoantigens including nucleic acids, histones, hnRNPs, snRNPs, collagens, Ro, La, SCL-70, CENP-B, Jo-1, pyruvate dehydrogenase, and post-translationally-modified antigens. Array analysis of serum derived from patients with SLE, rheumatoid arthritis, Sjogren's syndrome, mixed connective tissue diseases, scleroderma, myositidies, and primary biliary cirrhosis identified Autoantibody response patterns characteristic of these diseases (Fig. (Fig.1).1). We are now developing 'synovial proteome' arrays to study the Autoantibody responses in RA, and have produced first-generation arrays containing 450 distinct protein and peptide candidate antigens derived from synovial joints. These include native and citrulline-modified filaggrin peptides, native and deiminated fibrinogen and vimentin, glucose-6-phosphate isomerase, collagen types I, II, III, IV and V, hnRNP-A2/RA33, hnRNP-D, immunoglobulins, HCgp39, BiP, and HSPs 60, 65, 70, and 90. We probe our arrays with sera and synovial fluid from RA and control patients. We are using our arrays to examine serial serum samples from patients with early RA to: (1) identify antigens targeted early in disease, and (2) examine for evidence of B-cell epitope spreading. 'Synovial proteome' arrays represent a powerful tool to study the breadth and specificity of autoreactive B-cell responses, and to identify candidate and define relevant autoantigens in RA. Figure 1
K M Pollard - One of the best experts on this subject based on the ideXlab platform.
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advances in b cell epitope analysis of autoantigens in connective tissue diseases
Clinical Immunology, 2003Co-Authors: Michael Mahler, Martin Blüthner, K M PollardAbstract:The Characterization of Autoantibody specificities in rheumatic diseases is important in both diagnostic and basic research areas. Identification of the epitopes recognized by autoantibodies and their clinical and biological significance is not a trivial task. Epitopes may range in complexity from simple linear sequences of amino acids to complex quaternary structures. In addition to this structural complexity the frequency with which an autoantigen and its epitopes are recognized in a patient population may be useful in diagnosis, defining disease subgroups, and may offer information on disease prognosis. In this review recent advances in the epitope mapping of autoantigens in connective tissue diseases are discussed, with particular emphasis placed on the methodologies used to identify epitopes and the classification of the structural features of epitopes. To illustrate the identification of epitope structure, clinically relevant autoantigens, including CENP-A, PM/Scl-100, fibrillarin, filaggrin, Ro-52, and dsDNA, are discussed as examples of each type of epitope.
Rönnelid Johan - One of the best experts on this subject based on the ideXlab platform.
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Increased levels of anti-dsDNA antibodies in immune complexes before treatment with belimumab associate with clinical response in patients with systemic lupus erythematosus
'Springer Science and Business Media LLC', 2019Co-Authors: Sohrabian Azita, Parodis Ioannis, Carlströmer-berthén Nellie, Frodlund Martina, Jönsen Andreas, Zickert Agneta, Sjöwall Christopher, Bengtsson, Anders A., Gunnarsson Iva, Rönnelid JohanAbstract:Introduction: Immune complexes are of importance in systemic lupus erythematosus pathogenesis, and autoantibodies are believed to participate in immune complex formation. Quantification of Autoantibody levels in circulating IC might be of prognostic value. Methods: A C1q-binding-eluting technique was applied to purify immune complexes from 55 belimumab-treated systemic lupus erythematosus patients during a 24-month follow-up. Autoantibodies in serum and in solubilized immune complexes were quantified using addressable laser bead immunoassay. We investigated whether levels of autoantibodies in immune complexes associate with disease activity and response to belimumab treatment. Results: High baseline anti-double-stranded DNA and anti-histone levels in immune complexes associated with attainment of zero scores in clinical systemic lupus erythematosus disease activity index 2000 during the 24-month follow-up (p = 0.003 and p = 0.048, respectively). Low complement levels associated with high serum anti-double-stranded DNA and anti-ribosomal P levels (p = 0.003 and p = 0.008, respectively) and high anti-double-stranded DNA (p = 0.002) but not anti-ribosomal P levels in immune complexes. Anti-SSA/SSB serum levels were lower in patients attaining lupus low disease activity state at month 6; these associations were stronger for corresponding immune complex levels. Serum levels of most autoantibodies had declined at month 3, whereas Autoantibody levels in immune complexes, except for anti-double-stranded DNA, showed a more gradual decline over 1-2 years. Serum anti-double-stranded DNA levels decreased in all patients irrespective of systemic lupus erythematosus disease activity index 2000=0 attainment, whereas immune complex levels decreased only in achievers. Conclusion: Immune complex levels of autoantibodies against double-stranded DNA and the SSA/SSB complex show more specific associations with treatment outcome compared with serum levels in belimumab-treated systemic lupus erythematosus patients. Characterization of Autoantibody content in circulating immune complexes could prove useful in treatment evaluation in systemic lupus erythematosus and other immune complex-associated diseases
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Increased levels of anti-dsDNA antibodies in immune complexes before treatment with belimumab associate with clinical response in patients with systemic lupus erythematosus
'Springer Science and Business Media LLC', 2019Co-Authors: Sohrabian Azita, Parodis Ioannis, Carlströmer-berthén Nellie, Frodlund Martina, Jönsen Andreas, Zickert Agneta, Sjöwall Christopher, Bengtsson, Anders A., Gunnarsson Iva, Rönnelid JohanAbstract:Introduction Immune complexes are of importance in systemic lupus erythematosus pathogenesis, and autoantibodies are believed to participate in immune complex formation. Quantification of Autoantibody levels in circulating IC might be of prognostic value. Methods A C1q-binding-eluting technique was applied to purify immune complexes from 55 belimumab-treated systemic lupus erythematosus patients during a 24-month follow-up. Autoantibodies in serum and in solubilized immune complexes were quantified using addressable laser bead immunoassay. We investigated whether levels of autoantibodies in immune complexes associate with disease activity and response to belimumab treatment. Results High baseline anti-double-stranded DNA and anti-histone levels in immune complexes associated with attainment of zero scores in clinical systemic lupus erythematosus disease activity index 2000 during the 24-month follow-up (p = 0.003 and p = 0.048, respectively). Low complement levels associated with high serum anti-double-stranded DNA and anti-ribosomal P levels (p = 0.003 and p = 0.008, respectively) and high anti-double-stranded DNA (p = 0.002) but not anti-ribosomal P levels in immune complexes. Anti-SSA/SSB serum levels were lower in patients attaining lupus low disease activity state at month 6; these associations were stronger for corresponding immune complex levels. Serum levels of most autoantibodies had declined at month 3, whereas Autoantibody levels in immune complexes, except for anti-double-stranded DNA, showed a more gradual decline over 1-2 years. Serum anti-double-stranded DNA levels decreased in all patients irrespective of systemic lupus erythematosus disease activity index 2000=0 attainment, whereas immune complex levels decreased only in achievers. Conclusion Immune complex levels of autoantibodies against double-stranded DNA and the SSA/SSB complex show more specific associations with treatment outcome compared with serum levels in belimumab-treated systemic lupus erythematosus patients. Characterization of Autoantibody content in circulating immune complexes could prove useful in treatment evaluation in systemic lupus erythematosus and other immune complex-associated diseases.Funding Agencies|Swedish Research CouncilSwedish Research Council [521-13-3377]; Swedish Rheumatism Association [R757921, R-844801]; King Gustav V 80-year foundation [FAI-2017-0373, FAI-2018-0504]; ALF by the Uppsala County Council; Region Ostergotland; Agnes and Mac Rudberg Foundation; Ulla and Roland Gustafsson Foundation [2019-12]; Professor Nanna Svartz Foundation [2018-00250]; Brunnberg Foundation
Michael Bachmann - One of the best experts on this subject based on the ideXlab platform.
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Sequential use of immunoblots for Characterization of Autoantibody specificities.
Methods in molecular biology (Clifton N.J.), 2009Co-Authors: Holger Bartsch, Michael BachmannAbstract:Sera of patients with systemic autoimmune diseases frequently contain autoantibodies to nuclear autoantigens. Immunoblotting of recombinant and native autoantigens is a commonly used technique for the identification and Characterization of Autoantibody specificities. Here, we describe an easy procedure that facilitates the comparison of antibody specificities by reusing the same immunoblot at least three times in order to detect an abundantly expressed autoantigen in total cellular extracts.