Wang Xi, Dong Ke, Long Min, Lin Fang, Gao Zhaowei, Wang Lin, Zhang Zhe, Chen Xi, Dai Ying, Wang Huiping, Zhang Huizhong
Department of Medical Laboratory and Research Center, Tangdu Hospital, Fourth Military Medical University, Xinsi Road No 569, 710038, Xi'an, Shaanxi, China.
Arch Virol. 2018 Jul;163(7):1851-1861. doi: 10.1007/s00705-018-3797-7. Epub 2018 Mar 27.
Enterovirus 71 (EV71) is the most frequently detected causative agent in hand, foot, and mouth disease (HFMD) and is a serious threat to public health in the Asia-Pacific region. Many EV71 vaccines are under development worldwide, and although both inactivated virus vaccines and virus-like particles (VLPs) are considered to be effective, the main focus has been on inactivated EV71vaccines. There have been very few studies on EV71 VLPs. In this study, using a strategy based on HIV gag VLPs, we constructed a gag-VP1 fusion gene to generate a recombinant baculovirus expressing the EV71 structural protein VP1 together with gag, which was then used to infect TN5 cells to form VLPs. The VLPs were characterized using transmission electron microscopy, electrophoresis and staining with Coomassie blue, and Western blotting. Mice immunized with gag-VP1 VLPs showed strong humoral and cellular immune responses. Finally, immunization of female mice with gag-VP1 VLPs provided effective protection of their newborn offspring against challenge with a lethal dose EV71. These results demonstrate a successful approach for producing EV71 VP1 VLPs based on the ability of HIV gag to self-assemble, thus providing a good foundation for producing high-titered anti-EV71 antibody by immunization with VLP-based gag EV71 VP1 protein.
肠道病毒71型(EV71)是手足口病(HFMD)中最常检测到的病原体,对亚太地区的公共卫生构成严重威胁。全球许多EV71疫苗正在研发中,尽管灭活病毒疫苗和病毒样颗粒(VLP)都被认为是有效的,但主要重点一直放在灭活EV71疫苗上。关于EV71 VLP的研究很少。在本研究中,我们采用基于HIV gag VLP的策略,构建了一个gag-VP1融合基因,以产生一种表达EV71结构蛋白VP1和gag的重组杆状病毒,然后用其感染TN5细胞以形成VLP。通过透射电子显微镜、电泳、考马斯亮蓝染色和蛋白质印迹对VLP进行了表征。用gag-VP1 VLP免疫的小鼠表现出强烈的体液免疫和细胞免疫反应。最后,用gag-VP1 VLP免疫雌性小鼠为其新生后代提供了针对致死剂量EV71攻击的有效保护。这些结果证明了一种基于HIV gag自组装能力生产EV71 VP1 VLP的成功方法,从而为通过用基于VLP的gag EV71 VP1蛋白免疫产生高滴度抗EV71抗体奠定了良好基础。