Development of Reverse Genetics for the Prototype New World Mammarenavirus Tacaribe Virus.
Development of Reverse Genetics for the Prototype New World Mammarenavirus Tacaribe Virus.
复制标题
新世界乳腺病毒 Tacaribe 病毒原型的反向遗传学开发。
DOI：
10.1128/jvi.01014-20
复制
发表时间：
2020
影响因子：
5.4
通讯作者：
Martínez-Sobrido,Luis
中科院分区：
文献类型：
--
作者：
Ye,Chengjin;delaTorre,JuanCarlos;Martínez-Sobrido,Luis
已从果蝠、蚊子和蜱虫中分离出新世界乳头状病毒塔卡瑞病毒（TCRV），而所有其他已知的新世界乳头状病毒均存在于啮齿动物中。TCRV与人类疾病没有联系，但它已被证明可以保护感染了新世界乳头状病毒Junín病毒（JUNV）的狨猴免受阿根廷出血热样疾病的侵害，这表明TCRV有可能作为治疗阿根廷出血热的减毒活疫苗。建立对TCRV基因组进行遗传操作的反向遗传系统将促进TCRV作为减毒活疫苗或疫苗载体的实施。在这项研究中，我们首次开发了反向遗传学方法来产生重组TCRV （rTCRV）。我们成功地从双链病毒mRNA （rTCRV/GFP）中拯救了野生型（WT） rTCRV（一种表达两个报告基因的三段TCRV [r3TCRV]）和表达单个报告基因的二段TCRV。这些反向遗传学方法是研究TCRV生物学和探索其作为治疗其他病毒感染的减毒活疫苗或疫苗载体的潜在用途的极好工具。值得注意的是，我们在病毒大(L)片段的非编码基因间区（IGR）发现了39个核苷酸（nt）缺失（Δ39），这是优化病毒增殖所必需的。因此，在L-IGR中含有39-nt缺失的rTCRV （rTCRV/Δ39）在培养细胞中表现出病毒适应度下降，这表明在L-IGR中使用这种缺失作为减毒TCRV的一种方法，以及潜在的其他乳头状病毒作为减毒活疫苗或疫苗载体的可行性。迄今为止，没有食品和药物管理局（FDA）批准的疫苗可用于对抗人类由乳头状病毒感染引起的出血热。治疗乳头状病毒感染仅限于标签外使用利巴韦林，这是部分有效的，并与显著的副作用相关。Tacaribe病毒（TCRV）是新世界乳头状病毒的原型成员，对人类无致病性，但能够提供对阿根廷出血热病原体Junín病毒（JUNV）的保护，这表明使用TCRV作为治疗JUNV和潜在的其他病毒感染的减毒活疫苗载体的可行性。在这里，我们首次描述了利用反向遗传学方法产生重组TCRV （rTCRV）的可行性，这为研究TCRV的生物学以及它作为治疗人乳头状病毒和/或其他病毒感染的减毒活疫苗或疫苗载体的潜在用途铺平了道路。
The New World mammarenavirus Tacaribe virus (TCRV) has been isolated from fruit bats, mosquitoes, and ticks, whereas all other known New World mammarenaviruses are maintained in rodents. TCRV has not been linked to human disease, but it has been shown to protect against Argentine hemorrhagic fever-like disease in marmosets infected with the New World mammarenavirus Junín virus (JUNV), indicating the potential of TCRV as a live-attenuated vaccine for the treatment of Argentine hemorrhagic fever. Implementation of TCRV as a live-attenuated vaccine or a vaccine vector would be facilitated by the establishment of reverse genetics systems for the genetic manipulation of the TCRV genome. In this study, we developed, for the first time, reverse genetics approaches for the generation of recombinant TCRV (rTCRV). We successfully rescued a wild-type (WT) rTCRV (a trisegmented form of TCRV expressing two reporter genes [r3TCRV]) and a bisegmented TCRV expressing a single reporter gene from a bicistronic viral mRNA (rTCRV/GFP). These reverse genetics approaches represent an excellent tool to investigate the biology of TCRV and to explore its potential use as a live-attenuated vaccine or a vaccine vector for the treatment of other viral infections. Notably, we identified a 39-nucleotide (nt) deletion (Δ39) in the noncoding intergenic region (IGR) of the viral large (L) segment that is required for optimal virus multiplication. Accordingly, an rTCRV containing this 39-nt deletion in the L-IGR (rTCRV/Δ39) exhibited decreased viral fitness in cultured cells, suggesting the feasibility of using this deletion in the L-IGR as an approach to attenuate TCRV, and potentially other mammarenaviruses, for their implementation as live-attenuated vaccines or vaccine vectors.IMPORTANCETo date, no Food and Drug Administration (FDA)-approved vaccines are available to combat hemorrhagic fever caused by mammarenavirus infections in humans. Treatment of mammarenavirus infections is limited to the off-label use of ribavirin, which is partially effective and associated with significant side effects. Tacaribe virus (TCRV), the prototype member of the New World mammarenaviruses, is nonpathogenic in humans but able to provide protection against Junín virus (JUNV), the causative agent of Argentine hemorrhagic fever, demonstrating the feasibility of using TCRV as a live-attenuated vaccine vector for the treatment of JUNV and potentially other viral infections. Here, we describe for the first time the feasibility of generating recombinant TCRV (rTCRV) using reverse genetics approaches, which paves the way to study the biology of TCRV and also its potential use as a live-attenuated vaccine or a vaccine vector for the treatment of mammarenavirus and/or other viral infections in humans.