Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis
Multi-HLA class II tetramer analyses of citrulline-reactive T cells and early treatment response in rheumatoid arthritis
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DOI：
10.1186/s12865-020-00357-w
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发表时间：
2020-05-18
期刊：
影响因子：
3
通讯作者：
Malmstrom, Vivianne
中科院分区：
文献类型：
--
作者：
Gerstner, Christina;Turcinov, Sara;Malmstrom, Vivianne
背景HLA II类四聚体可用于抗原特异性CD 4 + T细胞的体外计数和表型表征。它们越来越多地应用于过敏，疫苗接种和自身免疫性疾病等环境。类风湿性关节炎（RA）是一种慢性自身免疫性疾病，其自身抗原种类繁多.结果利用多参数流式细胞术，我们开发了一种多HLA II类四聚体的方法，同时研究RA患者样本中的几种抗原特异性。我们重点关注先前描述的来自α-烯醇化酶、纤维蛋白原-β、波形蛋白以及软骨中间层蛋白（CILP）的瓜氨酸化HLA-DRB 1 *04：01限制性T细胞表位。首先，我们检查了来自健康供体（n = 7）的外周血中的测定间变异性和测定的灵敏度。接下来，我们在重复抽血的RA患者队列（n = 14）中证实了稳健性和灵敏度。然后，我们在两种不同的环境中应用我们的方法。我们评估了血清阳性关节痛（n = 5）和早期RA患者（n = 5）的淋巴组织，并在有发展RA风险的个体中显示出自身反应性T细胞。最后，我们研究了早期RA患者（n = 10）的外周血，发现在6个月随访时达到最低疾病活动度（DAS 28 < 2.6）的患者组显示瓜氨酸特异性T细胞频率降低。结论我们的研究表明，一个敏感的四聚体面板的发展，允许同时表征抗原特异性T细胞在体外患者样本，包括RA的“风险”的主题。这种多四聚体方法可用于在具有已知HLA限制性元件和几种候选抗原的任何疾病中进行纵向免疫监测。
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 alpha-enolase, fibrinogen-beta, 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.