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Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis

  • Gerstner, Christina1
  • Turcinov, Sara1
  • Hensvold, Aase H.1
  • Chemin, Karine1
  • Uchtenhagen, Hannes1, 2
  • Ramwadhdoebe, Tamara H.3, 4
  • Dubnovitsky, Anatoly1
  • Kozhukh, Genadiy1
  • Rönnblom, Lars5
  • Kwok, William W.2
  • Achour, Adnane6
  • Catrina, Anca I.1
  • van Baarsen, Lisa G. M.3, 4
  • Malmström, Vivianne1
  • 1 Karolinska University Hospital and Karolinska Institutet, Stockholm, Sweden , Stockholm (Sweden)
  • 2 BRI at Virginia Mason, Seattle, (WA), USA , Seattle (United States)
  • 3 Amsterdam UMC, University of Amsterdam, Amsterdam Infection & Immunity Institute, Amsterdam, Netherlands , Amsterdam (Netherlands)
  • 4 Academic Medical Center, Amsterdam, Netherlands , Amsterdam (Netherlands)
  • 5 Science for Life Laboratory, Uppsala, Sweden , Uppsala (Sweden)
  • 6 Karolinska University Hospital, Stockholm, Sweden , Stockholm (Sweden)
Published Article
BMC Immunology
Springer (Biomed Central Ltd.)
Publication Date
May 18, 2020
DOI: 10.1186/s12865-020-00357-w
Springer Nature


BackgroundHLA class II tetramers can be used for ex vivo enumeration and phenotypic characterisation of antigen-specific CD4+ T cells. They are increasingly applied in settings like allergy, vaccination and autoimmune diseases. Rheumatoid arthritis (RA) is a chronic autoimmune disorder for which many autoantigens have been described.ResultsUsing multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificities in RA patient samples. We focused on previously described citrullinated HLA-DRB1*04:01-restricted T cell epitopes from α-enolase, fibrinogen-β, vimentin as well as cartilage intermediate layer protein (CILP). First, we examined inter-assay variability and the sensitivity of the assay in peripheral blood from healthy donors (n = 7). Next, we confirmed the robustness and sensitivity in a cohort of RA patients with repeat blood draws (n = 14). We then applied our method in two different settings. We assessed lymphoid tissue from seropositive arthralgia (n = 5) and early RA patients (n = 5) and could demonstrate autoreactive T cells in individuals at risk of developing RA. Lastly, we studied peripheral blood from early RA patients (n = 10) and found that the group of patients achieving minimum disease activity (DAS28 < 2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells.ConclusionsOur study demonstrates the development of a sensitive tetramer panel allowing simultaneous characterisation of antigen-specific T cells in ex vivo patient samples including RA ‘at risk’ subjects. This multi-tetramer approach can be useful for longitudinal immune-monitoring in any disease with known HLA-restriction element and several candidate antigens.

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