负载重组口蹄疫病毒VP4蛋白的树突状细胞启动的T细胞应答

T Cell-mediated Responses Initiated by Dendritic Cells Pulsed with Recombinant VP4 Protein of Foot-and-mouth Disease Virus

为研究负载口蹄疫病毒VP4蛋白的树突状细胞对淋巴结T细胞的活化效应,构建了pET32a-VP4原核表达载体,经过诱导表达和纯化获得重组VP4蛋白。同时制备骨髓源树突状细胞(BMDCs)和淋巴结T细胞,以VP4蛋白负载BMDCs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法检测其IFN-γ的含量。结果显示,负载VP4蛋白的BMDCs与T细胞共培养后3、9和48h,试验组上清液中IFN-γ含量与对照组相比,均有极显著差异。这表明负载口蹄疫病毒VP4蛋白的BMDCs可有效激活淋巴结T细胞,使其分泌大量IFN-γ。

This study was designed to seek the activation effect on T lymphocytes by dendritic cells pulsed with recombinant VP4 protein of foot-and-mouth disease virus （FMDV）. To this end, we have successfully constructed the prokaryotic expression vector of pET32a-VP4. The re- combinant VP4 protein was expressed with induction of IPTG. VP4 protein was purified by SDS- PAGE and electro-elution approaches and identified with Western blot. Bone morrow-derived dendritic cells （BMDCs） were generated in vitro and pulsed with purified VP4 protein. The VP4- loaded BMDCs were co-cultured with lymph node T cells. Culture supernatants were harvested at indicated time points and the levels of IFN-γ were assayed with ELISA. The results showed that the IFN-γ levels of test groups were higher than that of control groups significantly at 3, 9, and 48 hours after co-culture. These data indicate that BMDCs pulsed with FMDV VP4 protein are a- ble to efficiently stimulate T lymphocyte activation and consequently initiate Thl-like response with enhanced secretion of IFN-γ