中文摘要：

目的：建立有效的脐血CD4^＋CD45^＋调节性T（Treg）细胞体外培养体系,探讨其对复发性流产患者免疫细胞功能的抑制作用,为开发新的复发性流产免疫治疗方法奠定实验基础。方法：应用免疫磁珠分选法从脐血淋巴细胞中获得Treg细胞,体外进行长期培养、扩增,分别应用MTT法和51Cr释放法检测扩增后的Treg细胞对复发性流产患者效应T细胞和NK细胞功能的抑制作用;ELISA法检测Treg对复发性流产外周血Th1/Th2细胞因子的影响。结果：经过5、15、25天培养,Treg细胞的扩增倍数分别为（6.5±0.6）倍、（18.2±2.5）倍和（52.7±4.8）倍,Treg细胞的纯度分别为（93.2±3.4）%、（88.6±2.9）%和（81.9±3.3）%。培养25天后的脐血Treg细胞对复发性流产的CD4＋CD25-效应T细胞和NK细胞杀伤活性具有明显抑制作用,能使外周血中IFNγ-水平下降,IL-10水平升高。结论：培养扩增后Treg细胞能在体外有效抑制复发性流产患者外周血免疫细胞功能,并能使外周Th1型细胞因子水平下降,Th2型细胞因子水平升高。

英文摘要：

Objective：To establish a practical and convenient approach of CD4^＋CD45^＋ regulatory T cells expansion in vitro and investigate the effect on the immunity of recurrent spontaneous abortion.Methods：CD4^＋CD45^＋ regulatory T cells were isolated from cord blood by MACS and then expand in vitro.Frequency and phenotype of Treg cells was determined by FACS.The inhibitory effect of expanded on effector T cells and NK cells was tested by MTT and 51Cr releasing method,respectively.Results：The amplification multiples of Treg cells were 6.5±0.6 folds,18.2±2.5 folds,and 52.7±4.8 folds,and the purity of Treg cells were（93.2±3.4）%,（88.6±2.9）% and（81.9±3.3）%,respectively.It was shown by the inhibitory test that the expanded Treg cells could inhibit the proliferation of CD4＋CD25-T cells and the specific lysis of NK cells in peripheral blood of RSA.The expanded Treg cells could down-regulate the level of IFN-γ and up-regulate the level of IL-10 in peripheral blood of RSA.Conclusion：The expanded Treg cells could significantly inhibit the immunity of recurrent spontaneous abortion.