IL-2抑制小鼠CD4^+ CD62L^+ T细胞分化为Th17细胞  

IL-2 inhibits the differentiation of mouse CD4^+ CD62L^+ T cells into Th17

作　　者：周骏[1] 王婷[1] 姚瑞芹[1] 曲学彬[1] 

机构地区：[1]徐州医学院基础医学院细胞生物学与神经生物学教研室,江苏徐州221009

出　　处：《基础医学与临床》2016年第1期19-23,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81302519);江苏省自然科学基金(BK20130221);江苏省研究生培养创新工程(KYZZ14_0398)

摘　　要：目的研究IL-2对小鼠脾脏CD4+CD62L+T细胞在体外向Th17细胞分化的作用。方法免疫磁珠法分选C57BL/6小鼠脾脏CD4+CD62L+T细胞,于抗体包被的培养板中培养3 d,实验分为对照组和IL-2处理组。对照组为经典Th17诱导分化培养基,IL-2处理组在对照组基础上于培养体系中添加IL-2。CFSE染色检测细胞增殖,Annexin V-PI法检测细胞凋亡,ELISA检测培养上清中IL-17A的浓度,荧光定量PCR检测Rorγt mRNA的表达,流式细胞术检测CD4^+IL-17^+Th17的生成比例以及Rorγt的表达。结果磁珠分选的小鼠脾脏CD4+CD62L+nave T细胞纯度高于95%。与对照组相比,IL-2处理组细胞数目明显增多,增殖能力明显增强(P<0.05),细胞凋亡比例降低(P<0.05);IL-2处理组培养上清中IL-17A的浓度明显降低(P<0.05),且CD4+IL-17+细胞比例下降,其特异性转录因子Rorγt的表达水平也显著降低(P<0.05)。结论 IL-2在CD4+CD62L+T细胞分化为Th17的过程中,能够促进T细胞的增殖并且抑制Th17的分化。Objective To investigate the effect of IL-2 on the differentiation of mice spleen CD4＋CD62L＋T cells into Th17 in vitro. Methods CD4^＋CD62L^＋T cells from C57 BL /6 mice with MACS divided into two groups,control group and IL-2 treated group were cultured in anti-CD3 and anti-CD28 antibody coated plate for 3 days. Control group cells were cultured in classical Th17 conditional medium,while IL-2 was added into the conditional medium of IL-2 treated group. Cell proliferation,cell apoptosis,the concentration of cytokine IL-17 A,expression level of specific transcription fator Rorγt,and percentage of CD4^＋IL-17^＋Th17 were measured by CFSE fluorescent staining,Annexin V-PI,ELISA,qRT-PCR,and FACS,respectively. Results The purity of separated CD4^＋CD62L^＋nave T was more than 95%. Compared with control group,the proliferation was improved（ P〈0. 05）with a reducing apoptosis（ P〈0. 05） in IL-2 treated cells. Moreover,IL-2 treated cells produced significantly lower concentration of IL-17A（ P〈0. 05）,decreased expression level of Rorγt（ P〈0. 05）,and less CD4^＋IL-17^＋cells（ P〈0. 05） as compared with control group. Conclusions During the process of CD4＋CD62L＋T differentiating into Th17 in vitro,IL-2 facilitate T cell proliferation but inhibits the differentiation of Th17.

关 键 词：IL-2 TH17 增殖 分化 

分 类 号：R392[医药卫生—免疫学]