Guanarito病毒(GTOV)是委内瑞拉出血热的病原体。GTOV属于哺乳动物属病毒,Arenaviridae家族,已被美国疾病控制和预防中心列为A类生物恐怖主义剂。尽管是一个高度优先的特工,没有针对委内瑞拉出血热的疫苗和药物。GTOVS-26764,从非致命人类病例中分离出来,在Vero细胞中产生不明确的细胞病变效应(CPE),对研究和对策发展工作构成重大障碍。与原始GTOVS-26764相比,本研究中产生的Vero细胞适应的GTOVS-26764产生了清晰的CPE,并在Vero细胞中显示出快速生长和高产率。我们为GTOV开发了反向遗传学系统,以研究通过Vero细胞适应获得的氨基酸变化并导致病毒表型变化。结果表明,L蛋白中的E1497K负责产生清晰的噬斑以及增强的病毒RNA复制和转录效率。Vero细胞适应性GTOVS-26764,能够产生CPE,将使研究人员能够轻松地对GTOV进行中和测定和抗药物筛选。此外,开发的反向遗传学系统将加速疫苗和抗病毒药物的开发。IMPORTANCEGuanarito病毒(GTOV)是一种啮齿动物传播的病毒。GTOV引起发烧,虚脱,头痛,关节痛,咳嗽,喉咙痛,恶心,呕吐,腹泻,鼻出血,牙龈出血,月经过多,和人类的黑莲娜。致死率为23.1%或更高。Vero细胞适应的GTOVS-26764显示出明显的细胞病变效应(CPE),而亲本病毒在Vero细胞中显示不清楚的CPE。我们产生了一个反向遗传学系统来拯救重组GTOV,并发现L蛋白中的E1497K负责形成清晰的斑块以及增强的病毒RNA复制和转录效率。这种反向遗传系统将加速疫苗和抗病毒药物的开发,这项研究的发现有助于理解GTOVL作为RNA聚合酶的功能。
Guanarito virus (GTOV) is the causative agent of Venezuelan hemorrhagic fever. GTOV belongs to the genus Mammarenavirus, family
Arenaviridae and has been classified as a Category A bioterrorism agent by the United States Centers for Disease Control and Prevention. Despite being a high-priority agent, vaccines and drugs against Venezuelan hemorrhagic fever are not available. GTOV S-26764, isolated from a non-fatal human case, produces an unclear cytopathic effect (CPE) in Vero cells, posing a significant obstacle to research and countermeasure development efforts. Vero cell-adapted GTOV S-26764 generated in this study produced clear CPE and demonstrated rapid growth and high yield in Vero cells compared to the original GTOV S-26764. We developed a reverse genetics system for GTOV to study amino acid changes acquired through Vero cell adaptation and leading to virus phenotype changes. The results demonstrated that E1497K in the L protein was responsible for the production of clear plaques as well as enhanced viral RNA replication and transcription efficiency. Vero cell-adapted GTOV S-26764, capable of generating CPE, will allow researchers to easily perform neutralization assays and anti-drug screening against GTOV. Moreover, the developed reverse genetics system will accelerate vaccine and antiviral drug development.IMPORTANCEGuanarito virus (GTOV) is a rodent-borne virus. GTOV causes fever, prostration, headache, arthralgia, cough, sore throat, nausea, vomiting, diarrhea, epistaxis, bleeding gums, menorrhagia, and melena in humans. The lethality rate is 23.1% or higher. Vero cell-adapted GTOV S-26764 shows a clear cytopathic effect (CPE), whereas the parental virus shows unclear CPE in Vero cells. We generated a reverse genetics system to rescue recombinant GTOVs and found that E1497K in the L protein was responsible for the formation of clear plaques as well as enhanced viral RNA replication and transcription efficiency. This reverse genetic system will accelerate vaccine and antiviral drug developments, and the findings of this study contribute to the understanding of the function of GTOV L as an RNA polymerase.