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通过包含本土聚合酶 PB1 蛋白增强反向遗传衍生的猪源 H1N1 流感病毒种子疫苗的生长。

Enhancement of reverse genetics-derived swine-origin H1N1 influenza virus seed vaccine growth by inclusion of indigenous polymerase PB1 protein.

机构信息

Virology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathumthani 12120, Thailand.

出版信息

Virus Res. 2010 Jan;147(1):145-8. doi: 10.1016/j.virusres.2009.10.010. Epub 2009 Oct 30.

Abstract

The current pandemic of a novel swine-origin H1N1 influenza virus (S-OIV) highlighted the need to urgently develop vaccines that can be used in a rapid response against the pathogen. Reverse genetics has been employed as an alternative means for the generation of influenza seed vaccines. However, reassortant viruses containing 6 internal genes from A/PR/8/34 and the hemagglutinin (HA) and neuraminidase (NA) genes from S-OIV showed very slow growth characteristics, hampering the speed of vaccine production. Here, we showed that the reverse genetics-derived H1N1 could be rescued with sensible viral titer by replacing PB1 of A/PR/8/34 with that of S-OIV for plasmid transfection. The "5+3" reassortant viruses have shown higher growth rate after transfection compared to that of "6+2" reassortant. The difference between PB1 of S-OIV and that of A/PR/8/34 in terms of the enhancement of virus growth was possibly due to the augmentation of viral polymerase activity, but not the lack of functional PB1-F2. Furthermore, it was found that growth enhancement by PB1 was specific for reassortant harboring HA of S-OIV, suggesting that the slow growth property of S-OIV reassortant virus is possibly due to restrictions imposed by the HA gene.

摘要

当前一种新型猪源 H1N1 流感病毒(S-OIV)的流行凸显了迫切需要开发能够快速应对病原体的疫苗。反向遗传学已被用作生成流感种子疫苗的替代手段。然而,含有来自 A/PR/8/34 的 6 个内部基因以及来自 S-OIV 的血凝素(HA)和神经氨酸酶(NA)基因的重组病毒表现出非常缓慢的生长特性,阻碍了疫苗生产的速度。在这里,我们表明通过用 S-OIV 的 PB1 替换 A/PR/8/34 的 PB1 可以拯救具有合理病毒滴度的反向遗传学衍生的 H1N1 用于质粒转染。与“6+2”重组体相比,转染后的“5+3”重组体显示出更高的生长速率。S-OIV 和 A/PR/8/34 的 PB1 之间在增强病毒生长方面的差异可能是由于病毒聚合酶活性的增强,而不是 PB1-F2 缺乏功能。此外,发现 PB1 的生长增强作用是特定于携带 S-OIVHA 的重组体的,这表明 S-OIV 重组病毒的生长缓慢特性可能是由于 HA 基因的限制。

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