负载灭活口蹄疫病毒树突状细胞启动的CD4^+T细胞应答  

CD4^+ T lymphocyte response triggered by dendritic cells pulsed with inactivated foot-and-mouth disease virus

作　　者：张丽[1] 石玮[1] 张雷[1] 李杰[1] 李娜[1] 安鹏丽[1] 张悦[1] 王蓓[2] 孟明[2] 王家鑫[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]河北大学医学部,河北保定071000

出　　处：《中国兽医科学》2011年第7期712-716,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30972168)

摘　　要：为研究负载灭活口蹄疫病毒(iFMDV)树突状细胞对CD4+T细胞的活化效应,用重组粒细胞-巨噬细胞集落刺激因子(rmGM-CSF)和白介素-4(rmIL-4)将小鼠外周血单核细胞诱导分化成树突状细胞(MoDCs),以信号阻断法制备淋巴结CD4+T细胞,利用负载iFMDV的MoDCs与CD4+T细胞共培养,以iFMDV+MoDCs作为对照,用ELISA检测不同时间培养上清液中IFN-γ的含量。用溶酶体抑制剂处理MoDCs,2h后再负载iFMDV,并与CD4+T细胞共培养,对照组则用iFMDV+MoDCs+CD4+T细胞,用ELISA检测不同时间培养上清液中IFN-γ的含量。结果显示,在共培养后第9和48小时,CD4+T细胞组与对照组比较差异均极显著(P<0.01)。共培养后第24和36小时,CD4+T细胞组与对照组比较均有显著性差异(P<0.05)。溶酶体抑制剂处理后,共培养第9、24、36和48小时组与对照组比较,均有显著性差异(P<0.05)。这表明MoDCs可通过溶酶体-MHC-Ⅱ途径向CD4+T细胞提呈iFMDV抗原,并且活化的CD4+T细胞可分化成Th1细胞。To investigate the activation of CD4＋ T cells by dendritic cells pulsed with inactivated foot-and-mouth disease virus（iFMDV） in vitro,monocyte-derived dendritic cells（MoDCs） were generated by culturing murine peripheral blood monocytes with addition of rmGM-CSF and rmIL-4 in the medium.CD4＋ T cells were harvested by blocking of CD8＋T cells with anti-CD8 antibody in the lymph node T cell pool.MoDCs pulsed with iFMDV were co-cultured with CD4＋ T cells,while iFMDV pulsed-MoDCs used as control.The release of the IFN-γ was determined at different time points with ELISA.The MoDCs were treated with lysosome inhibitor for 2 h and then pulsed with iFMDV and co-cultured with CD4＋ T cells.iFMDV pulsed-MoDCs co-cultured with CD4＋ T cell was designated as control.The supernatant levels of the IFN-γ were determined at different time points by ELISA.The levels of IFN-γ in supernatant of CD4＋ T cell culture were significantly higher than that of the control at hour 9 and 48 post co-culture（P0.01）.The quantity of IFN-γ was higher from hour 24 to 36 than that of the control.After treatment with chloroquine,the levels of IFN-γ in supernatant were significantly lower than those of control groups at hour 9,24,36 and 48 post co-culture（P 0.05）.These data indicated that MoDCs could capture iFMDV and process it in lysosome.The resultant peptides were loaded onto MHC-Ⅱ molecules for presentation to CD4＋ T cells,and that stimulated CD4＋ T cell differentiation into Th1 subset.

关 键 词：口蹄疫病毒 树突状细胞 CD4+T细胞 Γ干扰素 

分 类 号：S852.659.6[农业科学—基础兽医学]