类风湿关节炎患者蛋白聚糖aggrecan G1区特异性T细胞诱导SCID小鼠产生膝关节炎的实验研究  

Induction of arthritis in SCID mice by transferring aggrecan G1-specific human T-cells from rheumatoid arthritis patients

作　　者：朱剑[1] 黄烽[1] 张一平[2] 

机构地区：[1]解放军总医院风湿科,北京100853 [2]加利福尼亚大学尔湾分校神经病学系

出　　处：《解放军医学杂志》2006年第8期787-789,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助项目(30271233;30370226)

摘　　要：目的探讨针对蛋白聚糖的自身免疫反应在类风湿关节炎(RA)发病中的作用。方法从人软骨细胞中克隆获得ag-grecanG1区的cDNA片段,应用杆状病毒表达系统获取重组人aggrecanG1区蛋白(rAG1)。用rAG1刺激来自RA患者关节液的单个核细胞,采用β-液闪计数仪对培养细胞进行放射性检测并计算刺激增长指数(SI),取SI超过3者为rAG1特异性T细胞。将取自3个人的rAG1特异性T细胞克隆腹腔注射至8只SCIDBeige小鼠,同时在小鼠左膝注射rAG1。于注射后第8天处死小鼠,取双膝关节行组织病理学检查,观察炎细胞浸润及软骨和骨糜烂情况。结果从RA患者关节液单个核细胞分离得到rAG1特异性T细胞,主要为CD4+CD8-T细胞,分泌Th1型细胞因子(γ-干扰素)。部分腹腔注射人rAG1特异性T细胞的小鼠左膝出现了关节滑膜单个核细胞浸润和早期软骨侵蚀改变。结论人特异性rAG1反应性T细胞在rAG1刺激下可向关节局部归巢而导致关节炎症反应,提示针对rAG1的自身免疫反应可能参与了RA的发病。Objective To investigate the role of proteoglycan autoimmunity in the pathogenesis of rheumatoid arthritis （RA）. Methods Human Aggrecan G1 domain cDNA was cloned from human cartilage, and recombinant aggrecan G1 domain （rAG1） protein was synthesized in a Beculovirus expression system. Synovial fluid monocytes from RA patients were stimulated with rAG1. Harvested cells＇ radioactivity was quantified in a β-scentillation counter and the stimulation index （SI） was calculated. Cells with SI over 3 were taken as rAG1 specific T cells. Three well-characterized AG1-specific T cell lines were injected intraperitoneally into 8 SCID Beige mice. At the same time, rAG1 was injected intraarticularly into left knees of these mice. On day 8, mice were sacrificed and histological examination of the knee joints was performed. Results Human rAG1 specific T cell lines were generated from synovial fluid of RA patients. Most of these cells were CD4^＋CD8^- T cells secreting Thl cytokine （interferon-γ）. A pronounced capsular and synovial infiltration of mononuclear cells with early synovitis and cartilage erosion was observed in some left knees of the mice treated with rAG1-specific T cells. Conclusion Human rG1-autoreactive T cells injected intraperitoneally have homed to left knee where its epitope rAG1 was injected, and they participated in the development of inflammatory arthropathy.

关 键 词：关节炎 类风湿 自身免疫 蛋白聚糖类 

分 类 号：R392.3[医药卫生—免疫学] R593.22[医药卫生—基础医学]