类风湿性关节炎病人关节滑膜液与外周血中G1特异性T细胞的特点

The isolation of autoreactive T cells specific to aggrecan G1 domain from rheumatoid rathritis patients

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。

Objective To elucidate whether autoreactive T cells to G1 are expressed in rheumatoid arthritis patients.Methods We analyzed the frequency of human G1 specific T cells in the peripheral blood of four rheumatoid arthritis patients and tried to establish G1 reactive T cell lines from these rheumatoid arthritis patients with split well technique.ResultsThe G1 specific T cells in PBL were detectable at the range of 36.2% in peripheral blood lymphocytes.We have also generated 13 G1 specific T lymphocyte lines from these patients.All these cells expressed fine specificity to human recombinant G1,but not to unrelated antigen.They expressed apan T cell marker CD3 and CD4 and selectively used the αβ T cell receptor. Moreover,most of lines expressed the Th 1 cytokine profile,characterized by interferon γ positivity and IL 4 negativity.No Th 2 type cell line was generated.ConclusionThese data provide strong evidence in favor of the presence of autoreactive T cells in the rheumatoid arthritis patients. [