siRNA对γδT细胞Eomes基因表达的抑制功能研究  

Inhibitory effect of siRNA transfection on the expression of Eomesodermin gene in human γδT cells

作　　者：汪洪涛[1] 葛晓松[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,蚌埠233030

出　　处：《现代免疫学》2010年第1期51-55,共5页Current Immunology

基　　金：安徽省自然科学基金资助项目(050430603);安徽省教育厅省高校省级重点实验室重点科研项目(2004sys008);蚌埠医学院科研项目(BY0826)

摘　　要：本文旨在探讨RNA干扰对人外周血γδT细胞Eomes基因的表达和产生IFN-γ的抑制效应。化学合成3对EomessiRNA,脂质体转染试剂介导siRNA转染到用结核杆菌(Mtb)抗原刺激的人外周血单个核细胞中(主要是γδT细胞),通过转染条件优化后,对转染了siRNA的γδT细胞采用流式细胞术(FCM)进行分选,利用RT-PCR检测转染细胞EomesmRNA表达抑制率,以FCM检测转染前后γδT细胞IFN-γ产生的情况。结果显示,有2对siRNA能有效抑制Eomes的mRNA表达,并对γδT细胞IFN-γ的产生起抑制作用。提示Eomes siRNA可特异地抑制γδT细胞Eomes基因表达,Eomes可能是调控γδT细胞产生IFN-γ的重要转录因子。In the present study,the inhibitory effect of RNA interference on the expression of Eomesodermin（Eomes） gene in the human γδT cells from peripheral blood was investigated and the interferon-γ（IFN-γ） production was detected.Three pairs of small interference RNA（siRNA） with specific sequense for Eomes was synthesized and transferred into human γδT cells activated by Mtb-Ag by lipofectamine reagent.Under the optimized condition of transfection,Eomes siRNA transfected γδT cells were sorted by FCM and the inhibitory effect was detected by RT-PCR 24 hours after transfection.After transfection,the percentages of IFN-γ producing cells of γδT cells were detected by flow cytometry.The results showed that two pairs of siRNA inhibited Eomes gene expression and significantly decreased the IFN-γ secretion of γδT cells.These results suggest that the Eomes siRNA may be employed to specifically inhibit the Eomes gene expression in γδT cells and the transcription factor Eomes plays a critical role in the IFN-γ secretion of γδT cells.

关 键 词：SIRNA ΓΔT细胞 Eomes基因 

分 类 号：R392.11[医药卫生—免疫学]