PKCθ在结核杆菌抗原刺激人γδT细胞NFκB活化中的作用

The role of PKCθ in NFκB activation of human γδT cells stimulated by Mycobacterium Tuberculosis antigen

目的初步探讨PKCθ在结核杆菌抗原(MtbAg)刺激γδT细胞转录因子NFκB活化中的作用。方法 MtbAg刺激健康人外周血单个核细胞(PBMC),获得γδT细胞优势扩增细胞群。PKCθ选择性阻断剂rottlerin(粗糠柴毒素)预处理,MtbAg刺激γδT细胞,通过凝胶电泳迁移率变动分析(EMSA)检测γδT细胞NFκB活性,流式细胞术(FCM)检测γδT细胞活化分子CD69的表达。结果 MtbAg刺激30min,γδT细胞NFκB活性逐渐增加,刺激60min时NFκB活性显著增强;rottlerin使Mt-bAg激发的γδT细胞NFκB活性明显减弱。Rottlerin可显著抑制MtbAg诱导的γδT细胞活化分子CD69的表达(P<0.01)。结论 PKCθ参与MtbAg诱导的γδT细胞转录因子NFκB活化。

This study aimed to investigate the role of PKCθ in the activation of NFκB of γδT cells stimulated by MtbAg.Peripheral blood mononuclear cells（PBMC） from normal human subjects were stimulated by MtbAg and γδT cells were predominantly generated.The γδT cells were pretreated with rottlerin,a specific inhibitor of PKCθ,and then stimulated by MtbAg.NFκB activity was detected by EMSA,while CD69 expression was analyzed by flow cytometry.The NFκB activity of γδT cells stimulated by MtbAg was markedly blocked by rottlerin.Moreover,the expression of CD69,activation marker of γδT cells,was significantly inhibited by rottlerin.In conclusion,PKCθ was required for NFκB activation induced by MtbAg in γδT cells.