妊娠诱导的调节性T细胞对naiveT细胞抑制作用的体外研究  

In vitro study of inhibitory effects of pregnancy-induced regulatory T cells on naive T cells prolifera- tion

作　　者：周芹[1,2] 项世龙[1] 李琦[1] 蔡洁茹[1] 杨浩[1,2] 姜虹[1] 寿张飞[1,2] 陈江华[1,2] 

机构地区：[1]浙江大学医学院附属第一医院肾病中心,浙江省肾脏疾病防治技术研究重点实验室,国家中医药管理局三级实验室“肾脏病免疫实验室”,杭州310003 [2]卫生部多器官联合移植研究重点实验室

出　　处：《中华微生物学和免疫学杂志》2013年第7期495-500,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30872389）;浙江省自然科学基金（Y207088）

摘　　要：目的研究调节性T细胞（Treg）及妊娠诱导的调节性T细胞（piTreg）对naiveT细胞增殖的体外抑制，评价其抑制率的差异。方法对孕12．5d（E12．5d）异基因交配孕鼠C57／B6（早）xBALB／c（6）的piTreg和未孕雌鼠C57／B6鼠的Treg进行流式细胞术检测，观察piTreg和Treg在CD4＋ T细胞中比例及其胞内Voxp3表达变化；采用羧基荧光素二醋酸盐琥珀酰亚胺酯（CFSE）标记的naiveT作为效应细胞和丝裂霉素C灭活的CD4-T细胞作为刺激细胞的单向混合淋巴细胞培养体系，比较piTreg和Treg对效应细胞的体外抑制强度。结果piTreg水平明显高于Treg（P〈0．001），且这些细胞均表达高水平的Foxp3；piTreg在体外实验中对naXveT细胞的抑制效应明显高于Treg（P〈0．006），并受其细胞数量的影响。结论妊娠诱导的调节性T细胞较来自于非孕鼠的Treg对naXveT细胞有更明显的抑制作用，这一现象很可能是由妊娠中父系抗原刺激诱导的CD4＋CD25＋ Treg活性差异造成。Objective To investigate the in vitro inhibitory effects of regulatory T cells （Treg） from unpregnant mice and pregnancy-induced regulatory T cells （piTreg） on the proliferation of naive T cells and their differences. Methods The numbers of piTreg cells from allogeneic pregnant mice （ C57/B6 femalexBALB/c male） on day 12.5 （E12.5d） of gestation and Treg cells from unpregnant C57/B6 mice were detected respectively by flow cytometry. The percentages of piTreg ceils and Treg ceils in CD4＋ T cells of age-matched female mice and their intracellular expression of Foxp3 were analyzed . The in vitro inhibitory effects of piTreg cells and Treg cells on the CFSE-labeled naive T ceils （effector cells） were compared in a one-way mixed lymphocyte culture system using mitomycin C-inactivated CD4- T cells as stimulator cells. Results The level of piTreg cells in splenic mononuclear cells was significantly higher than that of Treg cells （P〈0. 001 ） from normal mice. Foxp3 was highly expressed in both piTreg cells and Treg cells, howev- er slightly increased in piTreg cells. Moreover, piTreg cells had a significant stronger in vitro inhibitory effect on naXve T cells proliferation than that of Tregs cells （P〈0.006）, which was in a cell-dependent manner. Conclusion The present study suggests that the piTreg cells have a stronger inhibitory effect on naive T cell proliferation as compared with Terg cells from unpregnant mice, The differential activity of CD4＋ CD25＋ Treg might be mediated by the paternal antigens during pregnancy.

关 键 词：piTreg 分选 扩增 抑制 

分 类 号：R714[医药卫生—妇产科学]