Yamagata Yutaro, Muramoto Yukiko, Miyamoto Sho, Shindo Keiko, Nakano Masahiro, Noda Takeshi
Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Laboratory of Ultrastructural Virology, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, Japan.
Microbiol Immunol. 2019 May;63(5):164-171. doi: 10.1111/1348-0421.12681. Epub 2019 May 17.
Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus-based antivirals.
缺陷干扰（DI）流感病毒在一个基因片段中存在大片段缺失，该缺失会干扰感染性病毒的复制；因此，这类病毒具有抗病毒治疗的潜力。然而，由于DI病毒在没有感染性辅助病毒帮助的情况下无法自主复制，所以尚未产生未被辅助病毒污染的克隆DI病毒株。为克服这一问题，我们利用反向遗传学技术构建了带有PB2 DI基因的克隆DI病毒，使用稳定表达PB2蛋白的细胞系扩增该克隆DI病毒，并证实了其在体外干扰感染性病毒复制的能力。因此，我们的方法适用于获得纯克隆DI病毒，将有助于理解DI病毒的干扰机制，并可用于开发基于DI病毒的抗病毒药物。
