Wang Hong-tao, Ge Xiao-song, Xue Zhu-ping, Li Bai-qing
Department of Immunology, Bengbu Medical College, Bengbu 233030, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Jan;26(1):31-4.
To investigate the effect of siRNAs specific for T-bet and Eomesodermin (Eomes) in interferon-gamma (IFN-gamma) production of different human T cell subsets.
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 alphabeta T cells and Mtb-Ag activated gammadelta T cells. CD4(+), CD8(+) T and gammadelta 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-gamma production were determined by flow cytometry.
After two series of transfection, the siRNA-FAM(+) cells were about 50% in the transfected cells. The IFN-gamma(+) cells in CD4(+) T cells (50.20%) decreased in the cells transfected with T-bet siRNA (18.46%) but no obvious decrease was observed in the cells transfected with Eomes siRNA, whereas the IFN-gamma(+) 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-gamma producing cells in gammadelta T cells (76.52%) decreased in the cells transfected with T-bet siRNA (56.57%) or with Eomes siRNA (42.53%).
At the level of transcription factors, T-bet and Eomes are important transcription factors for the regulation of IFN-gamma production in CD4(+) and CD8(+) T cells, respectively. Both T-bet and Eomes may simultaneously be involved in the regulation of the IFN-gamma production of gammadeltaT cells.
研究针对T-bet和胚外中胚层决定因子（Eomes）的小干扰RNA（siRNA）对不同人T细胞亚群产生干扰素-γ（IFN-γ）的影响。
化学合成针对T-bet和Eomes基因的双链小干扰RNA（siRNA）序列，并将其转染至抗CD3单克隆抗体激活的αβ T细胞和结核分枝杆菌抗原（Mtb-Ag）激活的γδ T细胞。通过流式细胞术分选CD4(+)、CD8(+) T细胞和γδ T细胞，采用逆转录-聚合酶链反应（RT-PCR）技术检测T-bet和Eomes基因mRNA的表达。通过流式细胞术测定IFN-γ产生的变化。
经过两轮转染后，转染细胞中siRNA-FAM(+)细胞约占50%。用T-bet siRNA转染的细胞中，CD4(+) T细胞中的IFN-γ(+)细胞（50.20%）减少（18.46%），而用Eomes siRNA转染的细胞中未观察到明显减少；在用Eomes siRNA转染的细胞中，CD8(+) T细胞中的IFN-γ(+)细胞（76.51%）减少（25.37%），而用T-bet siRNA转染的细胞中未观察到明显减少。然而，在用T-bet siRNA（56.57%）或Eomes siRNA（42.53%）转染的细胞中，γδ T细胞中产生IFN-γ的细胞（76.52%）减少。
在转录因子水平，T-bet和Eomes分别是调节CD4(+)和CD8(+) T细胞产生IFN-γ的重要转录因子。T-bet和Eomes可能同时参与调节γδ T细胞的IFN-γ产生。
