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

畜牧兽医学报

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

安鹏丽，李娜，李丽敏，张丽，张雷，高云欢，王家鑫^*

（河北农业大学动物医学院，保定 071000）

收稿日期:2012-03-27 出版日期:2012-10-25 发布日期:2012-10-25
通讯作者: 王家鑫，教授，E-mail: wangjiaxin66@yahoo.com.cn
作者简介:安鹏丽（1986-），女，河北无极人，硕士生，主要从事抗原提呈机制的研究。安鹏丽和李娜为并列第一作者
基金资助:
国家自然科学基金资助项目（30972168）

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

AN Peng-li, LI Na, LI Li-min, ZHANG Li, ZHANG Lei, GAO Yun-huan, WANG Jia-xin^*

(College of Veterinary Medicine, Agricultural University of Hebei, Baoding 071000, China)

Received:2012-03-27 Online:2012-10-25 Published:2012-10-25

摘要/Abstract

摘要： 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48 h，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。

Abstract: 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 recombinant 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 able to efficiently stimulate T lymphocyte activation and consequently initiate Th1-like response with enhanced secretion of IFN-γ.

中图分类号:

S852.659.6

安鹏丽，李娜，李丽敏，张丽，张雷，高云欢，王家鑫. 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[J]. 畜牧兽医学报, doi: .

AN Peng-li, LI Na, LI Li-min, ZHANG Li, ZHANG Lei, GAO Yun-huan, WANG Jia-xin. T Cell-mediated Responses Initiated by Dendritic Cells Pulsed with Recombinant VP4 Protein of Foot-and-mouth Disease Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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