体外应用MCMV与PMA/Ionomycin联合诱导Th17细胞的研究  

In vitro co-induction of Th17 cells by using MCMV and PMA/Ionomycin

作　　者：李旭芳[1] 舒赛男[1] 刘兴楼[1] 刘玲玲[1] 郭珊[1] 王慧[1] 李革[1] 方峰[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科病毒实验室,武汉430030

出　　处：《现代免疫学》2011年第3期234-237,共4页Current Immunology

基　　金：教育部博士点基金资助项目(20090142110076)

摘　　要：探讨体外联合应用小鼠巨细胞病毒(MCMV)和佛波酯(PMA)/离子霉素(Ionomycin)诱导Th17细胞产生。分别使用MCMV、PMA/Ionomycin或联合MCMV、PMA/Ionomycin体外刺激BALB/c小鼠脾细胞,利用流式细胞仪检测脾细胞中CD4+IL-17+细胞的比例及平均荧光强度,同时应用ELISA技术检测培养液上清中IL-17蛋白浓度。结果显示:单独使用MCMV或PMA/Ionomycin均不能有效刺激Th17细胞增殖,CD4+IL-17+细胞比例较低;而联合应用MCMV和PMA/Ionomycin可以有效诱导脾细胞向Th17细胞分化,CD4+IL-17+细胞比例达到2.6%,且应用联合诱导方案后可见CD4+IL-17+细胞平均荧光强度(MFI)明显升高。此外,联合应用MCMV和PMA/Ionomycin刺激脾细胞后明显增加培养液上清中IL-17蛋白浓度。我们的研究结果可以为进一步探讨Th17细胞在CMV感染免疫中的作用奠定实验基础。To investigate the co-induction of Th17 cells in vitro by using MCMV and PMA/Ionomycin,the splenocytes of BALB/c mice were stimulated with MCMV or PMA/Ionomycin or both of MCMV and PMA/Ionomycin.The proportion of CD4＋IL-17＋ cells in splenocytes was analyzed by flow cytometry and mean fluorescent intensity was also investigated.Meanwhile,the level of IL-17 in culture supernatant was detected by ELISA.Our results showed that usage of MCMV alone or PMA/Ionomycin to stimulate spenocytes could not induce Th17 cell differentiation efficiently and very lower proportion of CD4＋IL-17＋ cells was found.However,the use of MCMV and PMA/Ionomycin could obviously induce Th17 cell differentiation and the proportion of CD4＋IL-17＋ cells reached up to 2.6%,while the use of MCMV and PMA/Ionomycin could also significantly increase the mean fluorescent intensity of CD4＋IL-17＋ cells.Furthermore,the use of both MCMV and PMA/Ionomycin could remarkably increase the level of IL-17 in culture supernatants.Our results can establish the experimental foundation for the further study on the relationship between Th17 cells and the CMV infection.

关 键 词：CMV PMA/Ionomycin IL-17 平均荧光强度 

分 类 号：R392.4[医药卫生—免疫学]