离体CD4^+CD25^+调节性T细胞的扩增和混合淋巴细胞反应研究

Study on Proliferation and Mixed Lymphocyte Reactions of CD4^+CD25^+ Regulatory T Cells in Vitro

为研究健康人外周血CD4+CD25+调节性T细胞(CD4+CD25+regulatory T cells,CD4+CD25+Tregs)在协同刺激信号作用下的扩增反应及与CD4+CD25-T细胞混合淋巴细胞反应,采用免疫磁珠法分离CD4+CD25+Tregs和CD4+CD25-T细胞,在抗CD3-mAbs和抗CD28-mAbs的刺激下行CD4+CD25+Tregs培养和CD4+CD25+Tregs+CD4+CD25-T细胞混合淋巴细胞培养72 h。然后加入CCK-8溶液孵育1 h,用酶标仪检测OD450值。结果为:CD4+CD25+Tregs组OD450值极显著性地低于CD4+CD25-T细胞组(P<0.01)。CD4+CD25+Tregs与CD4+CD25-T细胞混合组OD450值也极显著性地低于CD4+CD25-T细胞组(P<0.01)。CD4+CD25+Tregs表现出无反应性特征,还可抑制CD4+CD25-T细胞扩增,因此,CD4+CD25+Tregs不只是很惰性的免疫细胞,而是对保持免疫耐受发挥了积极的调节作用。

Objective：to investigate the proliferation and lymphocyte reactions mixed with CD4 ＋ CD25- T cells of CD4 ＋ CD25＋ regulatory T cells （ CD4＋ CD25 ＋ Tregs） from peripheral blood obtained from healthy volunteers, effeeted by costimulatory singals. Methods ： CIM ＋ CD25 ＋ Tregs and CD4 ＋ CD25 - T cells were i- solated with magnetic cells sorting separation, then CD4 ＋ CD25 ＋ Tregs and CD4 ＋ CD25 ＋ Tregs ＋ CD4 ＋ CD25 - T cells were cuhered for 72 hours, stimulated by anti - CD3 - mAbs and anti - CD28 - mAbs. After that the 2 cultures added with CCK -8 solutin were incubated for 1 hour. Then OD450 were detected with mieroplate reader. Results ： OD450 of CD4 ＋ CD25＋ Tregs group was both extremely lower than that of CD4 ＋ CD25- T eells group（P 〈0.01） and that ofCD4＋ CD25＋ Tregs＋ CD4＋ CIY25- T cells group（P 〈0. 01）. Conelution ： CD4 ＋ CD25 ＋ Tregs were characterized with anergy, not proliferating, and inhabiting the proliferation of CD4 ＋ CD25 - T cells. So CD4 ＋ CD25 ＋ Tregs are not just inert immune cells but may play an active regulatory role in the maintenance of immunologic tolerance.