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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 prepare donor antigen-specific CD4 + CD25 + regulatory T cells and identify the induced cells for their immunobiological function. Methods: BALB / cByJ mice were immunized with splenocytes of donor ICR mice via tail vein once a week for 3 weeks. At the fourth week, CD4 + CD25 + T cells and CD4 + CD25-T cells were immunized with MACS. Mixed lymphocyte reaction and ELISA assay of cell function. Results: In vitro co-culture experiments showed that splenocytes from immunized BALB / cByJ mice showed a low proliferative response. CD4 + CD25 + T cells were able to inhibit the proliferation of CD4 + CD25-T cells stimulated by alloantigen-stimulated na [iota] ve BALB / cByJ mice and secretion of interferon gamma (IFN- [gamma]). CONCLUSION: CD4 + CD25 + regulatory T cells sorted after donor-specific splenocytes infusion are antigen-specific and immunocompetent and immunosuppressive in vitro.