SOCS3过表达对铜绿假单胞菌慢性肺部感染小鼠Th17免疫反应的影响  

The influence of SOCS3 overexpression on Th17 response in a mouse model of chronic Pseudomonas aeruginosa lung infection

作　　者：丁凤鸣[1] 陈宇清[1] 廖若敏[1] 张雪[1] 费霞[1] 张旻[1] 张杏怡[1] 

机构地区：[1]上海交通大学附属第一人民医院呼吸科,200080

出　　处：《免疫学杂志》2017年第5期389-393,399,共6页Immunological Journal

基　　金：国家自然科学基金(81300005)

摘　　要：目的探讨铜绿假单胞菌慢性肺部感染小鼠CD4^+T淋巴细胞SOCS3过表达对Th17免疫反应的作用。方法构建铜绿假单胞菌慢性肺部感染小鼠模型,并分离培养小鼠脾脏的CD4^+T淋巴细胞。构建SOCS3慢病毒过表达载体。在体外通过慢病毒基因转染的方法实现CD4^+T淋巴细胞SOCS3基因过表达。对SOCS3过表达细胞给予IL-23刺激,采用Western blot检测细胞中的p-STAT3水平,real-time PCR检测细胞中的RORγt水平,流式细胞分析IL-17+细胞数量,ELISA检测细胞培养液中的IL-17水平。结果铜绿假单胞菌慢性肺部感染小鼠CD4^+T淋巴细胞SOCS3过表达后,IL-23刺激诱导的p-STAT3蛋白水平、RORγt mRNA水平、IL-17^+细胞数量以及培养液中的IL-17水平均较对照组明显降低。结论体外研究表明SOCS3过表达对铜绿假单胞菌感染小鼠的Th17免疫反应具有抑制作用。This study aimed to investigate the changes of Th17 response when SOCS3 was overexpressed in CD4^＋ T cells isolated from a mouse model of chronic Pseudomonas aeruginosa lung infection. A recombinant lentivirus carrying SOCS3 gene was constructed and transfected into CD4^＋ T cells isolated from the spleens of mouse model. The SOCS3 overexpression in cells was confirmed by Western blotting and real-time PCR. Then the CD4^＋ T cells were stimulated with IL-23, and the expression of p-STAT3 was examined by Western blotting. The mRNA level of RORγt was examined by real-time PCR. The number of IL-17^＋ cells was analyzed by flow cytometry, and the level of IL-17 was assessed by ELISA. Data showed that SOCS3 overexpression significantly downregulated the levels of p-STAT3 protein and RORγt m RNA in CD4^＋ T cell; the number of IL-17^＋ cells and the level of IL-17 protein was significantly decreased in SOCS3-overexpressing cells compared with controls. These results of the in vitro study indicated that the overexpression of SOCS3 could suppress the Th17 response in mice with chronic Pseudomonas aeruginosa lung infection.

关 键 词：SOCS3 铜绿假单胞菌 慢性肺部感染 TH17 中性粒细胞 

分 类 号：R563.1[医药卫生—呼吸系统]