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43. Kongress der Deutschen Gesellschaft für Rheumatologie, 29. Jahrestagung der Deutschen Gesellschaft für Orthopädische Rheumatologie, 25. Wissenschaftliche Jahrestagung der Gesellschaft für Kinder- und Jugendrheumatologie

02.-05. September 2015, Bremen

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Identification and characterization of citrullinated antigen-specific B cells in peripheral blood of patients with rheumatoid arthritis

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  • Priscilla Kerkman - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • Emeline Fabre - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • Ellen van der Voort - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • Yoann Rombouts - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • Dominique Baeten - Academic Medical Center/University of Amsterdam, Clinical Immunology and Rheumatology, Amsterdam, The Netherlands
  • Tom Huizinga - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • René Toes - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands
  • Hans Ulrich Scherer - Leiden University Medical Center, Department of Rheumatology, Leiden, The Netherlands

Deutsche Gesellschaft für Rheumatologie. Deutsche Gesellschaft für Orthopädische Rheumatologie. Gesellschaft für Kinder- und Jugendrheumatologie. 43. Kongress der Deutschen Gesellschaft für Rheumatologie (DGRh); 29. Jahrestagung der Deutschen Gesellschaft für Orthopädische Rheumatologie (DGORh); 25. wissenschaftliche Jahrestagung der Gesellschaft für Kinder- und Jugendrheumatologie (GKJR). Bremen, 02.-05.09.2015. Düsseldorf: German Medical Science GMS Publishing House; 2015. Doc52.08 - ER.07

doi: 10.3205/15dgrh049, urn:nbn:de:0183-15dgrh0497

Veröffentlicht: 1. September 2015

© 2015 Kerkman et al.
Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). Lizenz-Angaben siehe http://creativecommons.org/licenses/by/4.0/.

Gliederung

Text

Introduction: Immunity to citrullinated antigens is a hallmark of rheumatoid arthritis (RA). We set out to elucidate the biology of this disease-specific immune response by identifying and characterizing citrullinated antigen-specific B cells in peripheral blood of RA patients.

Methods: Differentially labelled streptavidin and extravidin tetramers were conjugated to biotinylated CCP2 or control antigens and used in flow cytometry to identify citrullinated antigen-specific B cells in peripheral blood. Tetramer-positive and -negative B cells were isolated by FACS followed by in vitro culture and analysis of culture supernatants for the presence of ACPA by ELISA. Cells were phenotypically characterized by flow cytometry.

Results: By combining differentially labelled CCP2 tetramers, we successfully separated citrullinated antigen-specific B cells from non-specific background signals. Isolated tetramer-positive B cells, but not tetramer-negative cells, produced large amounts of ACPA upon in vitro stimulation. Phenotypic analyses revealed that citrullinated antigen-specific B cells displayed markers of class-switched memory B cells and plasmablasts, whereas only few cells displayed a naïve phenotype. The frequency of tetramer-positive cells was high (up to 1/500 memory B cells with a median of 1/12,500 total B cells) and correlated with ACPA serum titres and spontaneous ACPA production in culture.

Conclusion: We developed a unique technology to identify and isolate citrullinated antigen-specific B cells from peripheral blood of RA patients. Most cells have a memory phenotype, express IgA or IgG and are present in relatively high frequencies. These data pave the path for a direct and detailed molecular characterization of ACPA-expressing B cells and could lead to the identification of novel therapeutic targets.