摘要
目的探讨丝裂原活化的蛋白激酶(MAPK)信号通路在B7H1-Ig诱导Ⅰ型调节性T细胞(Tr1)分化过程中的作用。方法以预包被而固相化的小鼠B7H1-Ig融合蛋白加抗CD3单克隆抗体(单抗)刺激新鲜分离的C57BL/6小鼠初始CD4+CD62L+T细胞,诱导其向Tr1细胞分化。Western blotting检测MAPK信号通路(ERK1/2、p38 MAPK、JNK)的活化状况。在B7H1-Ig开始刺激时分别加入ERK1/2、p38和JNK通路特异性抑制剂PD98059、SB203580和SP600125,分别采用ELISA法、混合淋巴细胞反应(MLR)、流式细胞术和Western blotting分析检测抑制MAPK信号对B7H1-Ig刺激的CD4+T细胞产生的细胞因子分泌格局、功能及Foxp3表达的影响。结果 B7H1-Ig联合抗CD3单抗激活并诱导一群IL-10+IFN-γ+IL-5+IL-4low/-IL-2low/-Foxp3-Tr1细胞,通过分泌抑制性细胞因子IL-10发挥免疫抑制功能。Western blotting检测结果显示B7H1-Ig可激活CD4+T细胞中的p38 MAPK信号通路,对ERK1/2和JNK信号通路无明显影响。抑制p38 MAPK活性,可使B7H1-Ig诱导的CD4+T细胞IL-10和IL-5的分泌减少、免疫抑制功能减弱,促进B7H1-Ig刺激的CD4+T细胞向CD25+Foxp3+调节性T细胞(Treg)分化。结论 p38 MAPK信号通路的活化或抑制是参与调控由B7H1-Ig诱导的Tr1细胞分化及其与CD4+CD25+Foxp3+Treg之间相互转化的重要分子机制。
Objective To investigate the effects of mitogen-activated protein kinase （MAPK） signaling pathway on B7HI- Ig fusion pr0tein-initiated differentiation of type 1 regulatory T cells （Trl）. Methods Fresh isolated naive CD4 ＋ CD62L ＋ T cells of C57BL/6 mice were stimulated with immobilized B7HI-Ig fusion protein plus anti-CD3 monoelonal antibody to induce Trl cells. The activities of three major MAPK （ERKI/2, p38MAPK, JNK） signaling pathways during the process of Trl cells differentiation were analyzed by Western blotting. In experiments, the specific inhibitors of ERK1/2, p38 and JNK （PD98059, SB203580 and SP600125） were added separately at the time of culture initiation, and the influences on cytokines secretion, immune function and Foxp3 expression of B7Hl-Ig-stimulated CD4 ＋T cells were detected respectively by ELISA, mixed lymphocyte reaction （MLR）, flow cytometry and Western blotting. Results B7Hl-Ig-plus-anti-CD3 stimulation activated and induced the generation of a subset of IL-10 ＋ IFN-γ ＋ IL-5 ＋ IL-4^（low/-） IL-2^（low/-） Foxp3- Trl cells, which played immunosuppressive roles via secreting IL-10. Western blotting indicated that B7HI-Ig activated the p38 MAPK signaling pathway in CD4 ~T cells, while had no significant effect on ERK1/2 and JNK signaling pathways. Blocking p38 MAPK activity could significantly inhibit the production of IL-10 and IL-5 induced by B7HI-Ig in CD4 ~ T ceils, weaken the immunosuppressive function of B7Hl-Ig-stimulated CD4 ＋T cells and promote them to differentiate into CD25 ＋ Foxp3 ＊ Tregs. Conclusion The activation or inhibition of p38 MAPK signaling pathway is one of the important molecule mechanisms to control B7Hl-Ig-induced Trl cells differentiation and transformation between Trl cells and CD4 ＋ CD25 ＋ Foxp3 ＋ Tregs.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2012年第11期1448-1454,1453-1454,共7页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(30772018,30972691)
南通市科技计划项目(BK2011019)
江苏高校优势学科建设工程资助项目~~
