类风湿性关节炎患者关节滑液和滑膜中γδT 细胞的体外建系及功能研究

Establishment and functional studies on γδT cell lines derived from the synovial fluids and membranes of patients with rheumatoid arthritis

目的 探究在类风湿性关节炎（ Ｒ Ａ） 患者的关节滑液和滑膜中选择性存在，且有数量升高的γδ Ｔ 细胞的免疫学功能，以阐述类风湿性关节炎的发病机理。方法 应用固相抗体扩增法，建立关节滑液、滑膜和外周血γδ Ｔ 细胞系，并在此基础上，探讨了γδ Ｔ 细胞的细胞表型、细胞毒活性、增殖活性和细胞因子分泌情况。结果 所获得的滑液、滑膜γδ Ｔ 细胞系，多数以 Ｖδ１ 亚型细胞占优势；呈现出 Ｉ Ｌ２ 和 Ｉ Ｌ１５ 依赖的细胞增殖应答和显著增强的细胞毒活性；γδ Ｔ 细胞对自体和异体滑膜细胞的细胞毒作用无显著差异； Ｍ．ｔｂ（ 结核分枝杆菌） 、 Ｍ．ｔｂ 蛋白等抗原及有丝分裂原的刺激不能有效的提高建系的γδ Ｔ 细胞的细胞毒作用或增殖活性； 与异体滑膜细胞共同培养可以促进滑液γδ Ｔ 细胞的增殖； 细胞因子 Ｉ Ｌ２ 、 Ｔ Ｎ Ｆα的分泌维持在较低的水平。结论 存在于局部关节中的γδ Ｔ 细胞可能通过对滑膜细胞的细胞毒作用参与类风湿性关节炎的发病机制。

Establishment and functional studies on γδT cell lines derived from the synovial fluids and membranes of patients with rheumatoid arthritis$$$$ SONG Weihua,ZHANG Sumei,HE Wei,et al.Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & School of Basic Medicine , Peking Union Medical College,Beijing 100005 【 Abstract 】 Objective γδT cells in synovial fluids and membranes from patients with rheumatoid arthritis (RA) have been found to exist selectively.Understanding the functions of these γδT cells is important for elucidating the pathogenesis of RA. Methods An immobilized antibody expansion method was used to establish the γδT cell lines from synovial fluids,membranes and peripheral blood.The phenotypes,proliferations,cytotoxicities and cytokines secretions of γδT cell lines derived from synovial fluids and membranes were studied in vitro. Results The majority of γδT cell lines expressed TCR with the product of Vδ1 gene.They showed IL2 and IL15 dependent proliferations and their cytotoxicities were significantly enhanced by these cytokines. Mitogen such as PHAP and antigen such as M.tb, M.tb protein had no effect on their proliferations and cytotoxicities.Proliferation of γδT cells derived from synovial fluids increased if the cells were cocultured with allogeneic synovial cells and the cytotoxicity of these cells was not significant different to autologous and allogeneic synovial cells.Secretions of IL2 and TNFα by γδT cells maintained at a low level. Conclusion γδT cells in the joints might play a role in the pathogenesis of RA via their cytotoxicities. 【 Subject words 】 γδT cells Cytokine Rheumatoid arthritis