A Vero-cell-adapted vaccine donor strain of influenza A virus generated by serial passages
A Vero-cell-adapted vaccine donor strain of influenza A virus generated by serial passages
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通过连续传代产生的适用于 Vero 细胞的甲型流感病毒疫苗供体株
DOI：
10.1016/j.vaccine.2014.11.007
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发表时间：
2015-01-03
期刊：
影响因子：
5.5
通讯作者：
Sun, Bing
中科院分区：
文献类型：
--
作者：
Hu, Weibin;Zhang, Hong;Sun, Bing
基于细胞培养的疫苗生产系统优选用于流感疫苗的大规模生产，并且对于产生针对高致病性甲型流感病毒的疫苗具有优势。Vero细胞已被广泛用于人类疫苗生产，这些细胞的安全性已得到很好的证明。然而，最常用的流感疫苗供体病毒A/波多黎各/8/1934（PR 8）病毒在Vero细胞中不能有效生长。因此，我们通过连续传代使PR 8病毒适应Vero细胞，并在20代后获得高生长株。对适应株的序列分析和病毒学检测表明，NP、PB 1、PA和NS 1四个病毒内部基因的突变足以使适应株适应。携带这些突变的重组病毒（PR 8 - 4 mut）显示出加速的病毒转运到细胞核和增加的RNP活性。重要的是，PR 8 - 4 mut可以作为骨架供体病毒，支持H7 N1、H9 N2和H5 N1禽流感病毒以及H1N1和H3 N2人流感病毒在Vero细胞中的生长，而不会改变其在鸡胚或小鼠中的致病性。因此，我们的工作描述了Vero适应的高产PR 8 - 4 mut病毒的产生，该病毒可能作为流感疫苗供体病毒的有希望的候选者。(C)2014爱思唯尔有限公司版权所有。
A cell culture-based vaccine production system is preferred for the large-scale production of influenza vaccines and has advantages for generating vaccines against highly pathogenic influenza A viruses. Vero cells have been widely used in human vaccine manufacturing, and the safety of these cells has been well demonstrated. However, the most commonly used influenza-vaccine donor virus, A/Puerto Rico/8/1934 (PR8) virus, does not grow efficiently in Vero cells. Therefore, we adapted the PR8 virus to Vero cells by continuous passaging, and a high-growth strain was obtained after 20 passages. Sequence analysis and virological assays of the adapted strain revealed that mutations in four viral internal genes (NP, PB1, PA and NS1) were sufficient for adaptation. The recombinant virus harboring these mutations (PR8-4mut) displayed accelerated viral transport into the nucleus and increased RNP activity. Importantly, the PR8-4mut could serve as a backbone donor virus to support the growth of the H7N1, H9N2 and H5N1 avian viruses and the H1N1 and H3N2 human viruses in Vero cells without changing its pathogenicity in either chicken embryos or mice. Thus, our work describes the generation of a Vero-adapted, high-yield PR8-4mut virus that may serve as a promising candidate for an influenza-vaccine donor virus. (C) 2014 Elsevier Ltd. All rights reserved.