脐血CD4^+CD45^+调节性T细胞体外扩增及其抑制复发性流产患者免疫细胞功能研究  

Amplification of cord blood CD4^+CD45^+ regulatory T cells in vitro and its inhibition to the immunity of recurrent spontaneous abortion

作　　者：秦卫兵[1] 崔蓉[1] 杨宁[1] 王一峰[1] 苗竹林[1] 韦相才[1] 欧汝强[1] 

机构地区：[1]广东省计划生育科学技术研究所,广州510600

出　　处：《中国免疫学杂志》2011年第7期625-629,共5页Chinese Journal of Immunology

基　　金：广东省科技计划项目(No.2008B03031193);国家自然科学基金项目(No.30901610)资助课题

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。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.

关 键 词：CD4^+CD45^+Treg 体外扩增 复发性流产 

分 类 号：R392.9[医药卫生—免疫学]