转录因子T-bet和Eomes对不同亚类人T细胞产生IFN-γ的调控作用  

Role of transcription factor T-bet and Eomes in IFN-γ secretion of different human T cell subsets

作　　者：汪洪涛[1,2] 葛晓松[1,2] 薛祝平[1] 李柏青[1,2] 

机构地区：[1]蚌埠医学院免疫学教研室 [2]安徽省感染与免疫重点实验室,安徽蚌埠233030

出　　处：《细胞与分子免疫学杂志》2010年第1期31-34,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：安徽省自然科学基金项目(050430603);安徽省教育厅省高校省级重点实验室重点科研项目(2004sys008)

摘　　要：目的:探讨转录因子T-bet和Eomes对不同亚类人T细胞产生IFN-γ的调控作用。方法:化学合成针对人T-bet和Eomes基因的小干扰RNA(siRNA),转染体外培养的CD3单克隆抗体(mAb)活化的αβT细胞和Mtb-Ag活化的γδT细胞,利用流式细胞仪分选纯化CD4+、CD8+T细胞和γδT细胞,半定量RT-PCR检测CD4+、CD8+T淋巴细胞中T-bet和EomesmRNA的表达变化,流式细胞术(FCM)检测转染前后IFN-γ产生的变化情况。结果:经过连续2次转染,siRNA-FAM+细胞可达50%。转染T-bet-siRNA后,CD4+T细胞IFN-γ+细胞率(18.46±6.86)%,比阴性对照组(50.20±5.91)%,明显降低(P<0.01),而CD8+T细胞转染T-bet-siRNA,IFN-γ+细胞率(74.18±9.33)%,与阴性对照组(76.51±6.49)%无统计学意义;转染Eomes-siRNA,CD4+T细胞IFN-γ+细胞率(50.92±6.78)%,与对照组无统计学意义;而CD8+T细胞IFN-γ+细胞率(25.37±7.11)%,比阴性对照组(76.51±6.49)%,明显降低(P<0.01)。MtbAT转染T-bet-siRNA和Eomes-siRNA后,γδT细胞IFN-γ+细胞率分别为(56.57±6.67)%和(42.53±5.13)%,比阴性对照组(76.52±2.58)%均明显降低(P<0.01)。结论:在转录因子水平,T-bet和Eomes分别是调控CD4+、CD8+T淋巴细胞IFN-γ产生的重要转录因子,而两者可能同时参与了γδT细胞IFN-γ产生的调节。AIM：To investigate the effect of siRNAs specific for T-bet and Eomesodermin（Eomes）in interferon-γ（IFN-γ）production of different human T cell subsets.METHODS：Double-stranded small interfering RNA（siRNA）sequences specific for genes of T-bet and Eomes were chemically synthesized and transfected into anti-CD3 mAb activated αβ T cells and Mtb-Ag activated γδ T cells.CD4^＋,CD8^＋ T and γδ T cells were sorted by flow cytometry and the expressions of T-bet and Eomes gene mRNA were detected by RT-PCR technique.The changes of IFN-γ production were determined by flow cytometry.RESULTS：After two series of transfection,the siRNA-FAM＋ cells were about 50% in the transfected cells.The IFN-γ＋ cells in CD4^＋ T cells（50.20%）decreased in the cells trasnsfected with T-bet siRNA（18.46%）but no obvious decrease was observed in the cells trasnsfected with Eomes siRNA,whereas the IFN-γ＋ cells in CD8^＋ T cells（76.51%）decreased in the cells transfected with Eomes siRNA（25.37%）and no obvious decrease was observed in the cells transfected with T-bet siRNA.However,the IFN-γ producing cells in γδ T cells（76.52%）decreased in the cells transfected with T-bet siRNA（56.57%）or with Eomes siRNA（42.53%）.CONCLUSION：At the level of transcription factors,T-bet and Eomes are important transcription factors for the regulation of IFN-γ production in CD4^＋ and CD8^＋ T cells,respectively.Both T-bet and Eomes may simultaneously be involved in the regulation of the IFN-γ production of γδT cells.

关 键 词：SIRNA T-BET IFN-Γ 淋巴细胞 

分 类 号：R392.11[医药卫生—免疫学]