Arenavirus Genome Rearrangement for the Development of Live Attenuated Vaccines
Arenavirus Genome Rearrangement for the Development of Live Attenuated Vaccines
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DOI：
10.1128/jvi.00307-15
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发表时间：
2015-07-01
影响因子：
5.4
通讯作者：
Martinez-Sobrido, Luis
中科院分区：
文献类型：
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作者：
Cheng, Benson Yee Hin;Ortiz-Riano, Emilio;Martinez-Sobrido, Luis
沙粒病毒科的几个成员在人类中引起出血热疾病，并在其流行的地理区域造成严重的公共卫生问题以及可信的生物防御威胁。迄今为止，还没有FDA批准的沙粒病毒疫苗，目前的抗沙粒病毒治疗仅限于利巴韦林的标签外使用，仅部分有效。沙粒病毒是具有双节段负链RNA基因组的包膜病毒。每个基因组片段使用双义编码策略来指导两个方向相反的病毒多肽的合成，由非编码基因间区域分开。在这里，我们已经使用基于小基因组的方法来评估报告基因的表达水平，从核蛋白（NP）和糖蛋白前体（GPC）基因座内的原型沙粒病毒淋巴细胞性脉络丛脑膜炎病毒（LCMV）的S段。我们发现，报告基因表达更高的水平，从NP比从GPC位点。NP和GPC位点的报告基因表达水平的差异与重组三节段LCM病毒确认。然后，我们使用反向遗传学来拯救含有易位病毒S区段（rLCMV/TransS）的重组LCMV（rLCMV），其中病毒NP和GPC开放阅读框彼此替换。rLCMV/transS在培养的细胞中显示出较慢的生长动力学，并且在致死性LCMV感染的小鼠模型中在体内高度减毒，但是用rLCMV/transS免疫赋予针对野生型LCMV的致死性挑战的完全保护。rLCMV/ TransS的减毒与NP表达水平降低相关。这些结果为基于病毒基因组重排的沙粒病毒减毒活疫苗的开发开辟了一条新的途径。重要信息几种沙粒病毒可引起人类严重的出血热，并构成可靠的生物恐怖主义威胁。目前，没有FDA许可的疫苗可用于对抗沙粒病毒感染，抗沙粒病毒治疗仅限于利巴韦林的标签外使用，其仅部分有效且与副作用相关。在这里，我们描述了，第一次，产生的重组LCMV的病毒蛋白产品编码的S RNA片段（NP和GPC）被交换，以产生rLCMV/transS。rLCMV/TransS在培养细胞中表现出减少的病毒增殖，并且在体内高度减毒，同时在单次免疫剂量时赋予针对野生型LCMV的致死性攻击的保护。我们的研究为合理开发基于基因组重组的安全和保护性减毒活疫苗候选物提供了概念证明，用于治疗人类致病性沙粒病毒感染。
Several members of the Arenaviridae family cause hemorrhagic fever disease in humans and pose serious public health problems in their geographic regions of endemicity as well as a credible biodefense threat. To date, there have been no FDA-approved arenavirus vaccines, and current antiarenaviral therapy is limited to an off-label use of ribavirin that is only partially effective. Arenaviruses are enveloped viruses with a bisegmented negative-stranded RNA genome. Each genome segment uses an ambisense coding strategy to direct the synthesis of two viral polypeptides in opposite orientations, separated by a noncoding intergenic region. Here we have used minigenome-based approaches to evaluate expression levels of reporter genes from the nucleoprotein (NP) and glycoprotein precursor (GPC) loci within the S segment of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV). We found that reporter genes are expressed to higher levels from the NP than from the GPC locus. Differences in reporter gene expression levels from the NP and GPC loci were confirmed with recombinant trisegmented LCM viruses. We then used reverse genetics to rescue a recombinant LCMV (rLCMV) containing a translocated viral S segment (rLCMV/TransS), where the viral NP and GPC open reading frames replaced one another. The rLCMV/TransS showed slower growth kinetics in cultured cells and was highly attenuated in vivo in a mouse model of lethal LCMV infection, but immunization with rLCMV/TransS conferred complete protection against a lethal challenge with wild-type LCMV. Attenuation of rLCMV/ TransS was associated with reduced NP expression levels. These results open a new avenue for the development of arenavirus live attenuated vaccines based on rearrangement of their viral genome.IMPORTANCESeveral arenaviruses cause severe hemorrhagic fever in humans and also pose a credible bioterrorism threat. Currently, no FDAlicensed vaccines are available to combat arenavirus infections and antiarenaviral therapy is limited to the off-label use of ribavirin, which is only partially effective and associated with side effects. Here we describe, for the first time, the generation of a recombinant LCMV where the viral protein products encoded by the S RNA segment (NP and GPC) were swapped to generate rLCMV/TransS. rLCMV/TransS exhibited reduced viral multiplication in cultured cells and was highly attenuated in vivo while conferring protection, upon a single immunization dose, against a lethal challenge with wild-type LCMV. Our studies provide a proof of concept for the rational development of safe and protective live attenuated vaccine candidates based on genome reorganization for the treatment of pathogenic arenavirus infections in humans.