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


ABSTRACT: BACKGROUND:HLA 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. RESULTS:Using 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 ?-enolase, fibrinogen-?, 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?

SUBMITTER: Gerstner C 

PROVIDER: S-EPMC7236297 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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<h4>Background</h4>HLA 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.<h4>Results</h4>Using multi-parameter flow cytometry, we developed a multi-HLA class II tetramer approach to simultaneously study several antigen specificit  ...[more]

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