Wang Man, Jiang Shuai, Han Zhenwei, Zhao Bing, Wang Li'ao, Zhou Zhixia, Wang Yefu
Institute for Translational Medicine, Medical College of Qingdao University, Qingdao, 266021, China.
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
Appl Microbiol Biotechnol. 2016 Feb;100(3):1221-1230. doi: 10.1007/s00253-015-7027-x. Epub 2015 Oct 3.
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that is linked to the development of various malignancies. There is an urgent need for effective vaccines against EBV. EBV envelope glycoprotein gp350 is an attractive candidate for a prophylactic vaccine. This study was undertaken to produce the truncated (codons 1-443) gp350 protein (gp350(1-443)) in Pichia pastoris and evaluate its immunogenicity. The gp350(1-443) protein was expressed as a secretory protein with an N-terminal His-tag in P. pastoris and purified through Ni-NTA chromatography. Immunization with the recombinant gp350(1-443) could elicit high levels of gp350(1-443)-specific antibodies in mice. Moreover, gp350(1-443)-immunized mice developed strong lymphoproliferative and Th1/Th2 cytokine responses. Furthermore, the recombinant gp350(1-443) could stimulate CD4(+) and CD8(+) T cell responses in vaccinated mice. Collectively, these findings demonstrated that the yeast-expressed gp350(1-443) retained strong immunogenicity. This study will provide a useful source for developing EBV subunit vaccine candidates.
爱泼斯坦-巴尔病毒（EBV）是一种普遍存在的人类疱疹病毒，与多种恶性肿瘤的发生有关。迫切需要有效的抗EBV疫苗。EBV包膜糖蛋白gp350是预防性疫苗的一个有吸引力的候选物。本研究旨在利用毕赤酵母生产截短的（第1至443位密码子）gp350蛋白（gp350(1-443)）并评估其免疫原性。gp350(1-443)蛋白在毕赤酵母中作为一种带有N端His标签的分泌蛋白表达，并通过镍-亚氨基二乙酸（Ni-NTA）层析法纯化。用重组gp350(1-443)免疫可在小鼠中引发高水平的gp350(1-443)特异性抗体。此外，用gp350(1-443)免疫的小鼠产生了强烈的淋巴细胞增殖和Th1/Th2细胞因子反应。此外，重组gp350(1-443)可刺激接种疫苗小鼠的CD4(+)和CD8(+) T细胞反应。总体而言，这些发现表明酵母表达的gp350(1-443)保留了很强的免疫原性。本研究将为开发EBV亚单位疫苗候选物提供有用的资源。
