类风湿关节炎患者外周血与病变部位自身反应性T细胞的免疫学特征  

Activation and clonal expansion of T cells in the synovial fluid and peripheral circulation in patients with rheumatoid arthritis

作　　者：沈佰华[1] 王利[1] 张继英[1] 聂红[1] 余奇文[1] 陈广洁[1] 程琳玲[1] 柏峻[1] 李宁丽[1] 张冬青[1] 冯国璋 戴号 夏青 胡冬仪 许荣 倪立清 

机构地区：[1]上海第二医科大学上海市免疫学研究所 [2]上海市光华中西医结合医院

出　　处：《中华风湿病学杂志》2004年第8期458-462,共5页Chinese Journal of Rheumatology

基　　金：国家高技术发展计划(863)基金资助项目(2002AA216121);上海市科委发展基金资助项目(01JC14036;03DJ14009);上海市教委发展基金资助项目(03BZ02;03BK19)

摘　　要：目的研究类风湿关节炎(RA)外周血与病变部位自身反应性T细胞亚群的差异性。方法分析了35例RA患者滑膜液中与外周血T细胞的免疫表型、表面活化标志、Ⅱ型胶原和热休克蛋白70(HSP70)的反应频率与V茁的限制性取用。用流式细胞术分别测定滑膜液和外周血淋巴细胞表型和活化标志;采用标准3H-TdR掺入法分析了关节滑膜液中浸润的淋巴细胞对Ⅱ型胶原(CⅡ)和HSP70的反应频率,用半定量聚合酶链反应(PCR)检测了RA患者的V茁的限制性取用。结果滑膜液中CD4+T细胞与外周血中CD4+T细胞百分比相同,分别为40.0%和41.0%,滑膜液中CD8+T细胞比例升高,CD4/CD8细胞比值显著低于外周血。同时滑膜液中表达CD3和CD25的活化T细胞占(16±6)%,比外周血高。这群CD3/CD25双阳性T细胞对CⅡ和HSP70的反应频率增高。虽然外周血和滑膜液中T细胞受体均以琢茁TCR为主(70±16)%。但是进一步用流式细胞术分型检测其BV亚家族的取用特征,发现滑膜液中T细胞受体表现出限制性取用V茁14、16和17。结论在RA滑膜液中T细胞亚群明显不同于外周血;表现为活化细胞增多,对自身抗原的反应频率增高及T细胞受体的限制性取用。Objective To explore the difference between T cells in the synovial fluid and peripheral blood in patients with rheumatoid arthritis(RA). Method Samples from 22 patients were studied. The differentiation and activation markers expressed on T cell surface were detected by immunofluorscence using flow cytometer. The specific proliferation of collagen Ⅱ and heat shock protein 70 was analyzed using standard 3H-TdR incorporation method. Restricted V beta usage of these T cell was analyzed by semi-quantitied RT-PCR. Results The majority of the T cell subsets in the synovial fluid were demonstrated to be CD4 and CD8 positive cells in which (40±10)% were CD4 positive and (36±16)% were CD8 T cells respectively. The ratio between CD4 and CD8 was much lower than that found in the PBL of RA patients. The percentage of CD3+/CD25+ T cells was (16±6)%. The specific proliferation of collagen Ⅱ and HSP70 to CD3+/CD25+ T cell was higher than that of CD3+/CD25+ negative T cells. The T cell receptor expressed on the T cells from both peripheral blood and synovial fluid were tested for αβ TCR (70±26)%. However, the T cells in the synovial fluid showed Vβ14,16 and 17 restriction. Conclusion The data here reported indicates that T cell subsets in the synovial fluid and peripheral blood circulation in patients with rheumatoid arthritis are different. The T cells in the synovial fluid demonstrates more activation and higher reactivation to collagen Ⅱ and HSP70. The TCR of T cells showes Vβ14,16 and 17 restriction.

关 键 词：类风湿关节炎 外周血 病变部位 免疫学特征 T细胞亚群 免疫表型 

分 类 号：R593.22[医药卫生—内科学]