Virology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathumthani 12120, Thailand.
Virus Res. 2010 Jan;147(1):145-8. doi: 10.1016/j.virusres.2009.10.010. Epub 2009 Oct 30.
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.
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。