作　　者： ; ; ; ; ; ;

机构地区： 河北农业大学动物科技学院

出　　处： 《中国免疫学杂志》 2011年第11期970-973,977,共5页

摘　　要： 目的:研究负载口蹄疫病毒VP1蛋白质的树突状细胞对淋巴结T细胞产生IFNγ-的影响。方法:构建pET32a-VP1原核表达载体,经IPTG诱导表达并纯化重组蛋白VP1。制备骨髓源树突状细胞(BMDC)和淋巴结T细胞,将纯化的VP1蛋白质负载BMDC后与淋巴结T细胞共培养,收集不同时间点的共培养上清液,用ELISA检测其IFNγ-的含量。结果:本实验成功构建了pET32a-VP1原核表达载体,并获得了VP1蛋白质。在负载VP1蛋白质的BMDC与T细胞共培养后,实验组各时间点上清液的IFNγ-含量均高于对照组(3小时除外),特别是在共培养后9、24和48小时,差异显著。结论:负载FMDV VP1蛋白质的BMDC可有效激活淋巴结T细胞,从而启动Th1细胞免疫应答,分泌大量IFNγ-。 Objective：To study the activation of T lymphocytes by bone morrow-derived dendritic cells（BMDC） pulsed with VP1 protein of foot-and-mouth disease virus（FMDV） in vitro.Methods：The prokaryotic expression vector of pET32a-VP1 was constructed and induced to express VP1 by IPTG.VP1 protein was purified by common SDS-PAGE and electroelution approach.BMDC pulsed with FMDV VP1 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.Results：The prokaryotic expression vector was successfully constructed and VP1 protein was expressed.After co-culture of lymph node T cells with BMDC pulsed with VP1 protein,the IFN-γ levels of test groups were higher than control groups with significant differences at 9 h,24 h,and 48 h but no significant difference at 3 h.Conclusion：The BMDC pulsed with FMDV VP1 protein are able to activate the T cells efficiently,resulting in Th1 response with vigorous release of IFN-γ.

关 键 词： 口蹄疫病毒 蛋白质 原核表达 树突状细胞 细胞

领　　域： [农业科学] [农业科学] [农业科学]