抗原特异性T细胞活化后Foxp3、CTLA-4及细胞因子mRNA表达水平的动态分析  

The dynamic analysis of Foxp3,CTLA-4 and cytokine mRNA expressions in activated antigen-specific T cells

作　　者：季宝菊[1] 刘静[1] 招倩倩[1] 黄莺[1] 周小兵[2] 史利云[3] 邵启祥[1] 

机构地区：[1]江苏大学基础医学与医学技术学院免疫学与免疫学检验系,镇江212013 [2]江苏大学附属医院普外科,镇江212001 [3]杭州师范大学基础医学部免疫学和微生物学教研室,310036

出　　处：《免疫学杂志》2010年第5期416-418,422,共4页Immunological Journal

基　　金：国家自然基金资助项目(30671984);江苏省自然基金(BK2008231);江苏大学科技创新团队(2008-018-02);江苏大学学生科研立项(06A098)

摘　　要：目的研究抗原特异性T细胞活化后,其所表达的Foxp3、CTLA-4和细胞因子的动态变化,为深入了解特异性免疫应答调控奠定基础。方法体外采用树突状细胞(DC)系D2SC/1作为抗原提呈细胞(APC)将OVA323-339直接提呈给OVA特异性TCR转基因小鼠DO11.10来源的CD4^+T细胞。采用CCK-8法测定细胞活化增殖情况,并应用Real time-PCR动态分析细胞因子IL-2、IFN-γ、IL-10、TGF-β,CTLA-4和Foxp3的mRNA表达水平。结果OVA特异性T细胞活化所需的最佳OVA剂量为2μmol/L,T细胞活化后IL-2、IFN-γ、IL-10和TGF-βmRNA表达水平最高峰值分别在8、4、2 h和8 h,Foxp3 mRNA在2 h时有一过性的表达峰,随后在24 h又有所升高,而CTLA-4 mRNA在静止时有较高水平的表达,但活化后迅速下调,其在24 h内无明显变化。结论初步了解了适应性免疫应答中Foxp3、CTLA-4和主要免疫调节相关的细胞因子表达趋势,为深入研究适应性免疫应答调控机制奠定初步基础。We analyzed the dynamic change of the mRNA expression of Foxp3,CTLA-4 and cytokines in activated antigen specific T cells for learning more mechanism about the specific immune response.The dendritic cell（DC） line D2SC/1 were used as antigen presenting cells（APC） to present OVA_（323-339） to OVA-specific CD4~＋T cells divided from DOl 1.10 mouse,an OVA specific TCR transgenic mouse.Cell Counting Kit-8（CCK-8） was used to validate the activated T cell proliferation,while Real time-PCR was performed to evaluate the dynamic change of IL-2,IFN-γ,IL-10,TGF-β,CTLA-4,and Foxp3 mRNA.We found that the optimal dose of the OVA_（323-339） for activating the OVA-specific T cells was 2μmol/L;the peak time of the mRNA expression of IL-2,IFN-γ,IL-10 and TGF-βwere 8,4,2,and 8 h respectively,and Foxp3 mRNA had a temple peak at 2 h and up-regulated again after 24 h.CTLA- 4 mRNA had a high level in un-stimulated T cells,but down-regulated rapidly when T cell activated and maintained the lower level for more than 24 h.The evaluation of the dynamic regulation of the mRNA expression of Foxp3,CTLA-4 and the cytokines may lead us to learn and understand more about the mechanism of adaptive immune response.

关 键 词：抗原特异性 CD4+T 适应性免疫应答 

分 类 号：R392.11[医药卫生—免疫学]