抗原特异性CD4^+CD25^+T细胞的制备及功能鉴定  

Induction and identification of allogeneic antigen-specific CD4^+CD25^+ regulatory T cells

作　　者：张梅[1] 徐书杭[1] 徐瑜[1] 茅晓东[1] 刘翠萍[1] 徐宽枫[1] 刘超[1] 

机构地区：[1]南京医科大学第一附属医院内分泌科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第11期1590-1593,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省政府医学重点学科项目基金(苏卫科教2001-34)

摘　　要：目的:制备供体抗原特异性CD4+CD25+调节性T细胞,并对诱导的细胞进行免疫生物学功能鉴定。方法:分离供体ICR小鼠的脾细胞经尾静脉免疫BALB/cByJ小鼠,每周1次,连续3周。第4周,体外免疫磁珠分离技术(MACS)分选被免疫小鼠CD4+CD25+T细胞和CD4+CD25-T细胞。混合淋巴细胞反应和ELISA检测细胞的功能。结果:体外共培养实验显示,被免疫BALB/cByJ小鼠的脾细胞呈低增殖反应。CD4+CD25+T细胞能够抑制同种异体抗原刺激的天然(na觙ve)BALB/cByJ小鼠CD4+CD25-T细胞的增殖反应和γ干扰素(IFN-γ)的分泌。结论:供体特异性脾细胞输注后分选的CD4+CD25+调节性T细胞是抗原特异性的,在体外具有免疫无能性和免疫抑制性。Objective：To induce allogeneic antigen-specific CD4＋CD25＋ regulatory T cells in vivo and identify the function of these cells.Methods：Allogeneic antigen-specific CD4＋CD25＋ regulatory T cells were generated by the addition of multiple intravenous injections of ICR mice splenocytes in vivo.Spleen cells were then pooled and enriched for CD4＋CD25＋ T cells and CD4＋CD25-T cells by negative and positive selection using MACS in vitro.Characterizations of allogeneic antigen-specific CD4＋CD25＋ regulatory T cells were tested by mixed lymphocyte reaction（MLR） consisting of BALB/cByJ splenocytes as responder and MMC-treated ICR splenocytes as stimulator.The abilities of these cells to respond to various stimulators and to suppress alloreactive T cells responses were tested in vitro.Results：In the MLR,allogeneic antigen-specific T cells displayed little proliferation with allogeneic-antigen existence.Furthermore,these cells inhibited the proliferative responses and IFN-γ production of naive CD4＋CD25-T cells to alloantigens in vitro.Conclusion：Donor-specific splenocyte transfusion-induced CD4＋CD25＋ T cells were allogeneic antigen-specific.They were anergic,and retained their ability to suppress antigen-driven responses of CD4＋CD25-T cells in vitro.

关 键 词：CD4+CD25+调节性T细胞 同种异体抗原 无能 抑制 Γ干扰素 

分 类 号：R392.4[医药卫生—免疫学]