负载口蹄疫病毒VP1-VP4融合蛋白质的树突状细胞对T细胞的活化效应  

T lymphocyte activation by dendritic cells pulsed with VP1-VP4 fusion protein of foot-and-mouth disease virus

作　　者：李娜[1] 李丽敏[1] 安鹏丽[1] 高云欢[1] 董昌海[1] 王家鑫[1] 

机构地区：[1]河北农业大学动物医学学院免疫学实验室,河北保定071000

出　　处：《中国兽医学报》2012年第6期818-822,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：为研究负载口蹄疫病毒VP1-VP4融合蛋白质的树突状细胞对淋巴结T细胞的活化效应,通过构建pET32a-VP1-VP4原核表达系统制备VP1-VP4融合蛋白。将纯化VP1-VP4融合蛋白负载骨髓源树突状细胞(BMDC)后与淋巴结T细胞共培养,用ELISA检测不同时间点的共培养上清液中IFN-γ的含量。结果表明,负载FMDVVP1-VP4融合蛋白后的BMDC可通过溶酶体-MHC-Ⅱ类分子途径有效地激活淋巴结T细胞,从而启动Th1细胞免疫应答,分泌大量IFN-γ。To study the activation of T lymphocytes by bone marrow derived dendritic cells （BMDC） pulsed with re- combinant VP1-VP4 fusion protein of foot-and-mouth disease virus （FMDV） in vitro. The prokaryotic expression vector of pET32a-VP1-VP4 was constructed and VP1-VP4 fusion protein was expressed and purified by common SDS-PAGE and electroelution approach. BMDC pulsed with FMDV VP1-VP4 fusion protein were co-cultured with lymph node T cells. Supernatants were harvested at indicated time points and the IFN-γ levels of supernatants were determined with ELISA. The data show that BMDC pulsed with FMDV VP1-VP4 fusion protein are able to degrade VP1-VP4 antigen in lysosome and present the resulting peptides by MHC-II molecule for activating the T cells effi- ciently,leading to Thl-like response characteristic of release of IFN-γ.

关 键 词：口蹄疫病毒 VP1-VP4融合蛋白质 原核表达 树突状细胞 T细胞 

分 类 号：S852.65[农业科学—基础兽医学]