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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。
This article aims to investigate the RNAi interference on human peripheral blood γδT cells Eomes gene expression and IFN-γ inhibitory effect. Three pairs of EomessiRNAs were chemically synthesized and transfected into the peripheral blood mononuclear cells (mainly γδT cells) stimulated by Mtb antigens through liposome transfection reagent. After optimizing the transfection conditions, TheγδT cells stained with siRNA were sorted by flow cytometry (FCM). The expression of Eomes mRNA in transfected cells was detected by RT-PCR. The production of γδT-IFN-γwas detected by FCM. The results showed that two pairs of siRNA could effectively inhibit the expression of Eomes mRNA and inhibit the production of IFN-γ of γδT cells. These results suggest that Eomes siRNA can specifically inhibit the expression of Eomes in γδT cells. Eomes may be an important transcription factor that regulatesγδT cells producing IFN-γ.