Multi-HLA classification II tetramer analyses from citrulline-activated T tissue and early procedures impulse during the rheumatoid arthritis symptoms

HLA class II tetramers are used for old boyfriend vivo enumeration and you can phenotypic characterisation out-of antigen-specific CD4+ T structure. They are much more applied during the settings such sensitivity, inoculation and you will autoimmune illness. Rheumatoid arthritis (RA) are a long-term autoimmune problems wherein of a lot autoantigens had been revealed.

Performance

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*-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 < 2.6) at 6 months follow-up displayed a decrease in the frequency of citrulline-specific T cells.

Findings

The study demonstrates the development of a sensitive and painful tetramer panel enabling multiple characterisation off antigen-certain T structure during the ex vivo diligent samples and additionally RA ‘at risk’ sufferers. Continue Reading