Human IgG Fc promotes expression, secretion and immunogenicity of enterovirus 71 VP1 protein.
Human IgG Fc promotes expression, secretion and immunogenicity of enterovirus 71 VP1 protein.
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人 Ig G Fc 促进肠道病毒 71 VP1 蛋白的表达、分泌和免疫原性
DOI：
10.7555/jbr.30.20140157
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发表时间：
2016-05
影响因子：
2.3
通讯作者：
Zhang C
中科院分区：
文献类型：
--
作者：
Xu J;Zhang C
肠病毒（EV71）可引起严重的神经系统疾病，但其潜在的发病机制尚不清楚。衣壳蛋白病毒蛋白1 （VP1）在EV71的致病性中起关键作用。VP1蛋白的高水平表达和分泌是其天然构象结构、功能和免疫原性的必要条件。在我们前期的研究中，构建并分析了wt-VP1、tPA-VP1、VP1-d、VP1- hfc和VP1- mfc 5种密码子优化的VP1 DNA疫苗。它们表达VP1蛋白，但VP1构建体的分泌水平和免疫原性差异有统计学意义（P<0.05）。在本研究中，我们进一步研究了这些构建体的蛋白水平，并确定所有这些构建体都表达VP1蛋白。通过加入tPA先导序列可提高分泌水平，通过将人IgG Fc （hFc）与VP1融合可进一步提高分泌水平。VP1-hFc在小鼠中表现出最强的免疫原性。此外，hFc结构域可用于纯化VP1-hFc蛋白，用于进一步的研究。
Enterovirus (EV71) can cause severe neurological diseases, but the underlying pathogenesis remains unclear. The capsid protein, viral protein 1 (VP1), plays a critical role in the pathogenicity of EV71. High level expression and secretion of VP1 protein are necessary for structure, function and immunogenicity in its natural conformation. In our previous studies, 5 codon-optimized VP1 DNA vaccines, including wt-VP1, tPA-VP1, VP1-d, VP1-hFc and VP1-mFc, were constructed and analyzed. They expressed VP1 protein, but the levels of secretion and immunogenicity of these VP1 constructs were significantly different (P<0.05). In this study, we further investigated the protein levels of these constructs and determined that all of these constructs expressed VP1 protein. The secretion level was increased by including a tPA leader sequence, which was further increased by fusing human IgG Fc (hFc) to VP1. VP1-hFc demonstrated the most potent immunogenicity in mice. Furthermore, hFc domain could be used to purify VP1-hFc protein for additional studies.