ESAT-6对人外周血γδT细胞IL-17的影响及其信号通路的研究  

Effect of ESAT-6 on expression of interleukin-17 in γδT cells and its mechanism of signaling pathway

作　　者：王娟[1] 杜发旺[1] 杜先智[1] 

机构地区：[1]重庆医科大学第二附属医院呼吸内科,重庆400010

出　　处：《中国免疫学杂志》2014年第1期48-51,56,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.30872261)

摘　　要：目的:观察ESAT-6对人外周血γδT细胞表达IL-17的影响,并对其信号转导机制进行初步探讨。方法:用Ficoll密度梯度离心法从健康人外周血中分离单个核细胞,培养3天后用流式细胞仪分离纯化γδT细胞,并用唑来磷酸及IL-2刺激扩增γδT细胞,12天后用ESAT-6刺激γδT细胞,并给予信号传导及转录激活因子3(STAT3)抑制剂S3I-201,设置未加入ESAT-6及不加任何抑制剂的对照组,用PCR法检测IL-17、STAT-3的基因转录水平,用ELISA法检测细胞培养液中IL-17的含量,用Western blot检测STAT3蛋白的表达。结果:ESAT-6能显著提高STAT3及IL-17的基因转录及蛋白表达(P<0.01),且S3I-201能显著抑制ESAT-6所致的γδT细胞IL-17mRNA的转录及淋巴因子的增加(P<0.01)。结论:ESAT-6能提高外周血γδT细胞IL-17、STAT3的转录及表达,ESAT-6增强γδT细胞IL-17表达的机制可能与STAT3信号通路有关。Objective：To study the effect of ESAT-6 on expression of interleukin-17 in γδT cells and explore its mechanism of signaling pathway. Methods： Peripheral blood mononuclear cells （PBMC） were collected from health human blood with the ficoll density gradient centrifugation methods, and the T cells were gained by the nylon wool column method, and then the γδT cells were separated from the T cells with Flow cytometry. The zoledronie acid （ZA） and interleukiu-2 （IL-2） were used to stimulate and expand the γδT cells. The γδT cells were treated and stimulated by ESAT-6 after 12 days cultured, and the inhibitor of STAT3 signaling pathway （ S3I-201 ） was administered, besides control groups without ESAT-6 and stimulator were established. The change of IL-17 and STAT3 were determined by methods of PCR, and the expression of STAT3 protein was analyzed by the method of Western blot, and then the change of IL-17 was tested by ELISA. Results： ESAT-6 could increased the expression of IL-17 and STAT3 （P 〈 0.01 ） , and the inhibitor of STAT3 （ S3I-201 ） decreased expression of IL-17 （ P 〈 0.01 ）. Conclusion ： ESAT-6 can enhance the expression of IL-17 produced by γδT cells, and the mechanism of which maybe concerned with STAT3 signaling pathway.

关 键 词：ΓΔT细胞 白细胞介素17 ESAT-6 STAT3 

分 类 号：R392[医药卫生—免疫学]