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首页> 美国卫生研究院文献>Springer Open Choice >Influenza virus intracellular replication dynamics release kinetics and particle morphology during propagation in MDCK cells
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Influenza virus intracellular replication dynamics release kinetics and particle morphology during propagation in MDCK cells

机译：流感病毒在MDCK细胞中繁殖期间的细胞内复制动力学释放动力学和颗粒形态

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Influenza viruses are respiratory pathogens and can cause severe disease. The best protection against influenza is provided by annual vaccination. These vaccines are produced in embryonated chicken eggs or using continuous animal cell lines. The latter processes are more flexible and scalable to meet the growing global demand. However, virus production in cell cultures is more expensive. Hence, further research is needed to make these processes more cost-effective and robust. We studied influenza virus replication dynamics to identify factors that limit the virus yield in adherent Madin-Darby canine kidney (MDCK) cells. The cell cycle stage of MDCK cells had no impact during early infection. Yet, our results showed that the influenza virus RNA synthesis levels out already 4 h post infection at a time when viral genome segments are exported from the nucleus. Nevertheless, virus release occurred at a constant rate in the following 16 h. Thereafter, the production of infectious viruses dramatically decreased, but cells continued to produce particles contributing to the hemagglutination (HA) titer. The majority of these particles from the late phase of infection were deformed or broken virus particles as well as large membranous structures decorated with viral surface proteins. These changes in particle characteristics and morphology need to be considered for the optimization of influenza virus production and vaccine purification steps. Moreover, our data suggest that in order to achieve higher cell-specific yields, a prolonged phase of viral RNA synthesis and/or a more efficient release of influenza virus particles is required.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-016-7542-4) contains supplementary material, which is available to authorized users.
机译：流感病毒是呼吸道病原体，可导致严重疾病。年度疫苗接种可提供最佳的流感防护。这些疫苗是在有胚的鸡蛋中或使用连续的动物细胞系生产的。后者的流程更加灵活和可扩展，可以满足不断增长的全球需求。然而，在细胞培养物中生产病毒更昂贵。因此，需要进一步研究以使这些过程更具成本效益和更强大。我们研究了流感病毒的复制动力学，以找出限制粘附的Madin-Darby犬肾（MDCK）细胞中病毒产量的因素。 MDCK细胞的细胞周期阶段在早期感染期间没有影响。然而，我们的结果表明，当病毒基因组片段从细胞核中输出时，流感病毒RNA的合成已经在感染后4小时达到平衡。但是，在接下来的16小时内，病毒的释放速度是恒定的。此后，传染性病毒的产生急剧减少，但细胞继续产生有助于血凝（HA）滴度的颗粒。这些来自感染后期的颗粒大多数是变形或破碎的病毒颗粒以及装饰有病毒表面蛋白的大型膜状结构。为了优化流感病毒的生产和疫苗纯化步骤，必须考虑颗粒特征和形态的这些变化。此外，我们的数据表明，为了获得更高的细胞特异性产量，需要延长病毒RNA合成的阶段和/或更有效地释放流感病毒颗粒。电子补充材料本文的在线版本（doi：10.1007 / s00253-016-7542-4）包含补充材料，授权用户可以使用。

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