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目的为比较研究类风湿性关节炎病人 T细胞库中是否存在 G1特异性 T淋巴细胞极其生物学特性。方法应用重组人软骨抗原凝集原 G1为抗原 (r G1) ,采用国际标准半有限稀释建立细胞株法 ,从类风湿性关节炎病人外周血中分离建立 G1特异性 T细胞并分析其生物学特性。结果从 4名类风湿性关节炎病人外周血中分别分离建立了 13株 G1特异性 T细胞株。分析比较了这些细胞株 ,发现病人的 G1特异性 T淋巴细胞出现频率为 36 .2 %。所有 T细胞株与 r G1及 WT和 PPD共同培养以观察其特异性反应性 ,结果发现所建立的 13株 T细胞株对人软骨抗原凝集素 G1区具有良好的特异性。进一步用r G1肽段分析 ,发现其中 5株主要对 G1C末端呈特异性反应。细胞表型分析发现 ,所有这些 T细胞株均表现为 CD3+、CD4+与αβ TCR阳性 ;细胞因子分泌类型以 TH1为主。结论 RA病人外周血与关节滑膜液中均有 G1特异性 T细胞。
The purpose of this study was to compare the biological characteristics of G1-specific T lymphocytes in the T cell pool of rheumatoid arthritis patients. Methods G1 antigen (r G1) was cloned by recombinant human chondrocyte antigen. The cell line was established by semi-limited international standard. G1-specific T cells were isolated from the peripheral blood of patients with rheumatoid arthritis and their biological characteristics were analyzed . Results Thirteen G1-specific T cell lines were isolated from the peripheral blood of four rheumatoid arthritis patients. These cell lines were analyzed and compared, and the frequency of G1-specific T lymphocytes was found to be 36.2%. All T cell lines were co-cultured with rG1 and WT and PPD to observe their specific reactivity. As a result, the 13 T cell lines established showed good specificity for the human chondrogenic lectin G1 region. Further analysis of the rG1 peptide revealed that five of them mainly reacted specifically to the G1C terminus. Cell phenotype analysis found that all of these T cell lines showed CD3 +, CD4 + and αβ TCR positive; type of cytokine secretion TH1-based. Conclusion There are G1-specific T cells in peripheral blood and synovial fluid of RA patients.