Virology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand.
PLoS One. 2019 Mar 6;14(3):e0212632. doi: 10.1371/journal.pone.0212632. eCollection 2019.
More recently emerging strains of porcine epidemic diarrhea virus (PEDV) cause severe diarrhea and especially high mortality rates in infected piglets, leading to substantial economic loss to worldwide swine industry. These outbreaks urgently call for updated and effective PEDV vaccines. Better understanding in PEDV biology and improvement in technological platforms for virus production can immensely assist and accelerate PEDV vaccine development. In this study, we explored the ability of PEDV nucleocapsid (N) protein in improving viral yields in cell culture systems. We demonstrated that PEDV N expression positively affected both recovery of PEDV from infectious clones and PEDV propagation in cell culture. Compared to Vero E6 cells, Vero E6 cells expressing PEDV N could accelerate growth of a slow-growing PEDV strain to higher peak titers by 12 hours or enhance the yield of a vaccine candidate strain by two orders of magnitude. Interestingly, PEDV N also slightly enhances replication of porcine reproductive and respiratory virus, a PEDV relative in the Nidovirales order. These results solidify the importance of N in PEDV recovery and propagation and suggest a potentially useful consideration in designing vaccine production platforms for PEDV or closely related pathogens.
最近出现的猪流行性腹泻病毒(PEDV)株可导致感染仔猪严重腹泻和高死亡率,给全球养猪业带来巨大经济损失。这些疫情迫切需要更新和有效的 PEDV 疫苗。更好地了解 PEDV 的生物学特性和改进病毒生产技术平台,可以极大地帮助和加速 PEDV 疫苗的开发。在本研究中,我们探讨了 PEDV 核衣壳(N)蛋白在提高细胞培养系统中病毒产量的能力。我们证明,PEDV N 的表达可显著影响从感染性克隆中回收 PEDV 和在细胞培养中增殖 PEDV。与 Vero E6 细胞相比,表达 PEDV N 的 Vero E6 细胞可将生长缓慢的 PEDV 株的生长速度提高 12 小时,达到更高的峰值滴度,或使疫苗候选株的产量提高两个数量级。有趣的是,PEDV N 还略微增强了冠状病毒目副黏病毒科的猪繁殖与呼吸综合征病毒的复制。这些结果证实了 N 在 PEDV 恢复和增殖中的重要性,并提示在设计 PEDV 或密切相关病原体的疫苗生产平台时,这是一个潜在的有用考虑因素。