骨髓间充质干细胞体外诱导免疫调节机制的探讨  

Study of the immunoregulation mechanism of bone mesenchymal stem cells in vitro

作　　者：杜杰静[1] 宋红丽[1,2] 沈中阳[2] 郭瑞雪[2] 吴本娟[2] 付楠楠[2] 

机构地区：[1]天津医科大学一中心临床学院重点实验室,天津300192 [2]天津市第一中心医院器官移植中心,天津300192

出　　处：《天津医科大学学报》2012年第3期289-291,298,共4页Journal of Tianjin Medical University

基　　金：天津市应用基础及前沿技术重点研究计划项目(12JCZD-JC25200);天津市应用基础及前沿技术重点研究计划项目(11JCZDJC27800);天津市卫生局科技基金面上项目(2011KY11)

摘　　要：目的:观察骨髓间充质干细胞(BM MSCs)对体外活化T细胞的免疫调节作用。方法:提取Wistar大鼠BM MSCs及新鲜脾淋巴细胞做共培养。实验分3组,3个时间点:分别为0、24、48 h脾淋巴细胞/BM MSCs+刀豆蛋白A(ConA)(实验组),脾淋巴细胞+ConA(对照组),单纯淋巴细胞组(空白组)。MTT检测T淋巴细胞活性,ELISA检测细胞因子TNF-α、IL-10及TGF-β水平,流式检测Foxp3+调节性T细胞表达率。结果:(1)与对照组相比,实验组24、48 h在2个时间点T淋巴细胞活性均显著降低(P<0.05);(2)与对照组相比,实验组TNF-α在2个时间点有显著降低(P<0.05),实验组IL-10、TGF-β表达水平有显著升高(P<0.05);(3)实验组Foxp3+T调节细胞表达率高于对照组;(4)随着时间延长,空白组及对照组T淋巴细胞活性均显著降低(P<0.05),实验组T淋巴细胞活性随时间延长差异无统计学意义。结论:BM MSCs可以抑制T淋巴细胞活化,并通过减少炎症因子TNF-α的释放抑制免疫应答。另外,BM MSCs可通过自分泌及诱导T调节细胞产生并分泌免疫抑制因子IL-10、TGF-β下调免疫反应,产生免疫耐受。Objective： To investigate the immunoregulation mechanism of allogeneic bone mesenchymal stem cells（BM MSCs） on T lymphocyte cell proliferation.Methods： Several BM MSCs and T lymphocyte cell from Wistar rats were cocultured.There were 3 groups,comprising splenic lymphocyte/BM MSCs＋ConA（experimental group）,splenic lymphocyte＋ConA（control group）,splenic lymphocyte（blank）.Detect the viability of T lymphocyte by MTT,detect the level of cell factor TNF-α,IL-10 and TGF-β by ELISA,and detect the expression percentage of Foxp3＋ regulatory T cells by flow cytometry.Results：（1）T lymphocyte cell proliferation were significantly lower in experimental group compared with control group（P0.05）.（2）TNF-α were significantly lower in experimental group compared with control group（P0.05）.IL-10,TGF-β were significantly greater in experimental group compared with control group（P0.05）.（3）The level of regulatory T cell in experimental group were significantly greater compared with control group（P0.05）.（4）With the extent of time,the level of T lymphocyte cells were significantly decreased.Conclusion： T lymphocyte cell proliferation are significantly lower in experimental group compared with control group,so is the level of TNF-α.But BM MSCs can secrete IL-10 and TGF-β and boost the level of regulatory T cell to promote immune tolerance.

关 键 词：骨髓间充质干细胞 T淋巴细胞 免疫调节 共培养 大鼠 

分 类 号：R392.1[医药卫生—免疫学]