负载灭活口蹄疫病毒树突状细胞启动的CD8^+T细胞应答  

CD8+ T lymphocyte response triggered by dendritic cells pulsed with inactivated foot-and-mouth disease virus

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

出　　处：《中国兽医学报》2012年第3期415-419,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972168)

摘　　要：探讨在体外条件下负载灭活口蹄疫病毒(iFMDV)树突状细胞对CD8+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),用信号阻断法从淋巴结T细胞制备CD8+T细胞,利用负载iFMDV的MoDCs与CD8+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。用蛋白酶体抑制剂处理MoDCs,2h后再负载iFMDV,并与CD8+T细胞共培养,对照组则用iFMDV+MoDCs+CD8+T细胞,用ELISA检测不同时间培养上清液中IFN-γ的含量。结果表明,同对照组相比,在共培养后9,12,24,36,48h,CD8+T细胞组均产生高水平的IFN-γ。而且,经抑制剂预处理的MoDCs与CD8+T细胞共培养后,其分泌的IFN-γ量不仅没有降低,反而显著升高。因此得出,MoDCs可与蛋白酶体等加工iFMDV抗原,并通过交叉提呈途径激活CD8+T细胞。To explore the activation of CD8＋T cell by dendritic cells pulsed with inactivated foot-and-mouth disease virus（iFMDV） in vitro,monocyte-derived dendritic cells（MoDCs） were generated by culturing peripheral blood monoytes with addition of rmGM-CSF and rmIL-4 in the medium.CD8＋T cells were prepared by blocking of CD4＋T cells with anti-CD4 antibody in the lymph node T cell pool.MoDCs pulsed with iFMDV were co-cultured with CD8＋T cells.iFMDV pulsed-MoDCs were used as control.The release of IFN-γ was determined at indicated timepoints with ELISA.The MoDCs were treated with proteasome inhibitor for 2 h and pulsed with iFMDV for co-culture with CD8＋T cells.iFMDV pulsed-MoDCs co-cultured with CD8＋T cells was utilized as control.The level of IFN-γ in supernatant was determined at indicated timepoints by using ELISA.The levels of IFN-γ in supernatant of CD8＋T cell culture were higher than those of the control at 9 h,12 h,24 h,36 h,and 48 h post co-culture.The levels of IFN-γ released from CD8＋T cells triggered by iFMDV-pulsed MoDCs treated with lactacystin were higher than those of control group.These findings show that MoDCs have a capability of processing iFMDV in proteasome and other unidentified enzyme-containing compartments and corss-presenting antigenic peptides to CD8＋T cells.

关 键 词：口蹄疫病毒 树突状细胞 CD8+T细胞 Γ-干扰素 

分 类 号：S852.65[农业科学—基础兽医学] R535[农业科学—兽医学]