Development of Live-Attenuated Arenavirus Vaccines Based on Codon Deoptimization
Development of Live-Attenuated Arenavirus Vaccines Based on Codon Deoptimization
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DOI：
10.1128/jvi.03401-14
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发表时间：
2015-04-01
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
沙粒病毒对公共卫生有重大影响，并构成可信的生物防御威胁，但安全有效的沙粒病毒疫苗的开发仍然难以捉摸，目前没有食品和药物管理局（FDA）许可的沙粒病毒疫苗可用。在这里，我们探索了使用基于密码子去优化（CD）的方法作为开发减毒沙粒病毒活疫苗的新策略。我们重新编码的原型沙粒病毒淋巴细胞脉络丛脑膜炎病毒（LCMV）的核蛋白（NP）在哺乳动物细胞中使用频率最低的密码子，这导致较低的LCMV NP的表达水平在转染细胞中，与降低NP活性在基于细胞的功能测定。我们使用反向遗传学方法来拯救一组重组LCMV（rLCMV）编码CD NP（rLCMV/NPCD），其在体外显示减弱的生长动力学。此外，使用充分表征的LCMV感染小鼠模型的实验显示，rLCMV/NPCD 1和rLCMV/NPCD 2在体内高度减毒，但在单次免疫后，对随后的野生型（WT）重组LCMV（rLCMV/WT）致死性攻击提供完全保护。在FDA疫苗批准的Vero细胞中连续传代期间，rLCMV/NPCD 1和rLCMV/NPCD 2均具有遗传和表型稳定性。这些结果提供了一个基于CD的方法开发减毒活疫苗候选人对人类致病性arenavirus.IMPORTANCESeveral arenaviruses造成严重的出血热在人类和生物恐怖主义威胁的安全性，有效性和稳定性的概念证明。目前，没有FDA许可的疫苗可用于对抗沙粒病毒感染，而抗沙粒病毒治疗仅限于利巴韦林的标签外使用，这只是部分有效，并与副作用有关。在这里，我们描述了原型沙粒病毒LCMV编码密码子去优化病毒核蛋白（rLCMV/NPCD）的重组版本的产生。Weidentified rLCMV/NPCD 1和rLCMV/NPCD 2在体内高度减毒，但能够提供保护，防止随后的致命挑战与野生型LCMV。这些病毒在培养细胞中连续扩增传代期间显示减毒表型。我们的研究结果支持使用这种方法来开发安全，稳定和保护性的减毒沙粒病毒活疫苗。
Arenaviruses have a significant impact on public health and pose a credible biodefense threat, but the development of safe and effective arenavirus vaccines has remained elusive, and currently, no Food and Drug Administration (FDA)-licensed arenavirus vaccines are available. Here, we explored the use of a codon deoptimization (CD)-based approach as a novel strategy to develop live-attenuated arenavirus vaccines. We recoded the nucleoprotein (NP) of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) with the least frequently used codons in mammalian cells, which caused lower LCMV NP expression levels in transfected cells that correlated with decreased NP activity in cell-based functional assays. We used reverse-genetics approaches to rescue a battery of recombinant LCMVs (rLCMVs) encoding CD NPs (rLCMV/NPCD) that showed attenuated growth kinetics in vitro. Moreover, experiments using the well-characterized mouse model of LCMV infection revealed that rLCMV/NPCD1 and rLCMV/NPCD2 were highly attenuated in vivo but, upon a single immunization, conferred complete protection against a subsequent lethal challenge with wild-type (WT) recombinant LCMV (rLCMV/WT). Both rLCMV/NPCD1 and rLCMV/NPCD2 were genetically and phenotypically stable during serial passages in FDA vaccine-approved Vero cells. These results provide proof of concept of the safety, efficacy, and stability of a CD-based approach for developing live-attenuated vaccine candidates against human-pathogenic arenaviruses.IMPORTANCESeveral arenaviruses cause severe hemorrhagic fever in humans and pose a credible bioterrorism threat. Currently, no FDA-licensed vaccines are available to combat arenavirus infections, while antiarenaviral therapy is limited to the off-label use of ribavirin, which is only partially effective and is associated with side effects. Here, we describe the generation of recombinant versions of the prototypic arenavirusLCMVencoding codon-deoptimized viral nucleoproteins (rLCMV/NPCD). Weidentified rLCMV/NPCD1 and rLCMV/NPCD2 to be highly attenuated in vivo but able to confer protection against a subsequent lethal challenge with wild-type LCMV. These viruses displayed an attenuated phenotype during serial amplification passages in cultured cells. Our findings support the use of this approach for the development of safe, stable, and protective live-attenuated arenavirus vaccines.