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BMC Immunology
Published in: BMC Immunology 1/2020

Open Access 01-12-2020 | Joint Pain | Research article

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

Authors: Christina Gerstner, Sara Turcinov, Aase H. Hensvold, Karine Chemin, Hannes Uchtenhagen, Tamara H. Ramwadhdoebe, Anatoly Dubnovitsky, Genadiy Kozhukh, Lars Rönnblom, William W. Kwok, Adnane Achour, Anca I. Catrina, Lisa G. M. van Baarsen, Vivianne Malmström

Published in: BMC Immunology | Issue 1/2020

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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 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.

Conclusions

Our 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.
Appendix
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Metadata
Title
Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis
Authors
Christina Gerstner
Sara Turcinov
Aase H. Hensvold
Karine Chemin
Hannes Uchtenhagen
Tamara H. Ramwadhdoebe
Anatoly Dubnovitsky
Genadiy Kozhukh
Lars Rönnblom
William W. Kwok
Adnane Achour
Anca I. Catrina
Lisa G. M. van Baarsen
Vivianne Malmström
Publication date
01-12-2020
Publisher
BioMed Central
Published in
BMC Immunology / Issue 1/2020
Electronic ISSN: 1471-2172
DOI
https://doi.org/10.1186/s12865-020-00357-w

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