通过改变膜融合活性,博尔纳病病毒糖蛋白中的单个氨基酸替换可提高水泡性口炎病毒伪型病毒的感染滴度。

IF 1.9 4区 医学 Q4 IMMUNOLOGY Microbiology and Immunology Pub Date : 2024-09-23 DOI:10.1111/1348-0421.13172
Yusa Akiba, Hiromichi Matsugo, Takehiro Kanda, Modoka Sakai, Akiko Makino, Keizo Tomonaga
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引用次数: 0

摘要

博尔纳病病毒 1(BoDV-1)会导致包括人类在内的哺乳动物患上急性致命性脑炎。尽管博尔纳病病毒 1 非常重要,但由于其在细胞中产生的病毒颗粒感染性低,而且缺乏细胞病理效应(通常用于筛选),因此有关博尔纳病病毒 1 进入细胞的研究一直受到阻碍。为了解决这些问题,我们开发了一种方法来高效生产伪型为包括 BoDV-1 在内的正脊病毒属成员糖蛋白(G)的水泡性口炎病毒(VSV)。我们发现,要获得 VSV 假型病毒的高感染性滴度,必须有最佳的 G 表达。值得注意的是,带有来自 BoDV-1 株系 huP2br 的 G 的 VSV 假型病毒的感染率明显高于带有来自 He/80 株系的 G 的 VSV 假型病毒的感染率。突变分析表明,BoDV-1-G残基307上的蛋氨酸增加了以BoDV-1-G为假型的VSV(VSV-BoDV-1-G)的感染性滴度。细胞-细胞融合试验表明,该残基在膜融合中起着关键作用,从而表明高膜融合活性和膜融合的宽 pH 值范围是 VSV-BoDV-1-G 获得高感染性滴度的关键。我们的研究还有助于确定 BoDV-1-G 的功能域,并深入了解 G 介导的细胞进入。
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A single amino acid substitution in the Borna disease virus glycoprotein enhances the infectivity titer of vesicular stomatitis virus pseudotyped virus by altering membrane fusion activity

Borna disease virus 1 (BoDV-1) causes acute fatal encephalitis in mammals, including humans. Despite its importance, research on BoDV-1 cell entry has been hindered by low infectious viral particle production in cells and the lack of cytopathic effects, which are typically useful for screening. To address these issues, we developed a method to efficiently produce vesicular stomatitis virus (VSV) pseudotyped with glycoprotein (G) of members of the genus Orthobornavirus, including BoDV-1. We discovered that optimal G expression is required to obtain a high infectivity titer of the VSV pseudotyped virus. Remarkably, the infectivity of the VSV pseudotyped virus with G from the BoDV-1 strain huP2br was significantly higher than that of the VSV pseudotyped virus with G from the He/80 strain. Mutational analysis demonstrated that the methionine at BoDV-1–G residue 307 increases the infectivity titer of VSV pseudotyped with BoDV-1–G (VSV–BoDV-1–G). A cell‒cell fusion assay indicated that this residue plays a pivotal role in membrane fusion, thus suggesting that high membrane fusion activity and a broad pH range for membrane fusion are crucial for achieving a high infectivity titer of VSV–BoDV-1–G. This finding may be extended to increase the infectivity titer of VSV pseudotyped virus with other orthobornavirus G. Our study also contributes to identifying functional domains of BoDV-1–G and provides insight into G-mediated cell entry.

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来源期刊
Microbiology and Immunology
Microbiology and Immunology 医学-免疫学
CiteScore
5.20
自引率
3.80%
发文量
78
审稿时长
1 months
期刊介绍: Microbiology and Immunology is published in association with Japanese Society for Bacteriology, Japanese Society for Virology, and Japanese Society for Host Defense Research. It is peer-reviewed publication that provides insight into the study of microbes and the host immune, biological and physiological responses. Fields covered by Microbiology and Immunology include:Bacteriology|Virology|Immunology|pathogenic infections in human, animals and plants|pathogenicity and virulence factors such as microbial toxins and cell-surface components|factors involved in host defense, inflammation, development of vaccines|antimicrobial agents and drug resistance of microbes|genomics and proteomics.
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