A Lassa Fever Live-Attenuated Vaccine Based on Codon Deoptimization of the Viral Glycoprotein Gene
A Lassa Fever Live-Attenuated Vaccine Based on Codon Deoptimization of the Viral Glycoprotein Gene
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DOI：
10.1128/mbio.00039-20
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发表时间：
2020-01-01
期刊：
影响因子：
6.4
通讯作者：
Martinez-Sobrido, Luis
中科院分区：
文献类型：
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作者：
Cai, Yingyun;Ye, Chengjin;Martinez-Sobrido, Luis
拉萨病毒(LASV)在西非流行，估计每年感染数十万人。其中相当数量的感染导致拉沙热(LF)，在住院确诊患者中与显著的发病率和高达69%的病死率有关。美国食品和药物管理局批准的LF疫苗不可用。目前的抗病毒治疗仅限于标签外使用核苷类似物利巴韦林，该药物只有部分有效，并与显著的副作用有关。我们构建了表达密码子去优化(CD)糖蛋白前体基因(GPC)的重组LASV，rLASV-GPC/CD。生长动力学和峰值滴度的比较表明，与野生型(WT)重组LASV(rLASV-WT)相比，rLASV-GPC/CD在细胞培养中略有减弱。然而，rLASV-GPC/CD在13株和Hartley豚鼠中高度减弱，反映在接种rLASV-GPC/CD的动物中没有可检测到的临床症状。重要的是，单次皮下注射rLASV-GPC/CD可提供完全的保护，防止致命的LASV暴露。我们的结果证明了CD方法用于开发安全有效的LASV减毒活疫苗候选疫苗的可行性。此外，rLASV-GPC/CD可能为研究人员提供一种在最大(生物安全级别4)容器外安全研究LASV的工具，这将加速LASV分子和细胞生物学的基本方面的阐明和新LASV医学对策的开发。没有获得许可的LF疫苗，抗LF治疗仅限于在标签外使用核苷类似物利巴韦林，疗效不确定。我们描述了一种新的LASV减毒活疫苗候选疫苗的产生。该候选疫苗是基于病毒基因组关键区域的野生型(WT)LASV的突变，即糖蛋白前体(GPC)基因。这些突变不会改变编码的GPC，但会干扰其在宿主细胞中的产生。这种突变的LASV(rLASV-GPC/CD)在细胞培养中的表现与WT LASV(rLASV-WT)相似，但与rLASV-WT不同，在接种的豚鼠中不会引起疾病。用rLASV-GPC/CD免疫的豚鼠对WT LASV的致死性暴露有保护作用。我们的结果支持该候选疫苗在非人类灵长类动物LF模型中的测试。
Lassa virus (LASV) is endemic in Western Africa and is estimated to infect hundreds of thousands of individuals annually. A considerable number of these infections result in Lassa fever (LF), which is associated with significant morbidity and a case-fatality rate as high as 69% among hospitalized confirmed patients. U.S. Food and Drug Administration-approved LF vaccines are not available. Current anti-viral treatment is limited to off-label use of a nucleoside analogue, ribavirin, that is only partially effective and associated with significant side effects. We generated and characterized a recombinant LASV expressing a codon-deoptimized (CD) glycoprotein precursor gene (GPC), rLASV-GPC/CD. Comparison of growth kinetics and peak titers showed that rLASV-GPC/CD is slightly attenuated in cell culture compared to wild-type (WT) recombinant LASV (rLASV-WT). However, rLASV-GPC/CD is highly attenuated in strain 13 and Hartley guinea pigs, as reflected by the absence of detectable clinical signs in animals inoculated with rLASV-GPC/CD. Importantly, a single subcutaneous dose of rLASV-GPC/CD provides complete protection against an otherwise lethal exposure to LASV. Our results demonstrate the feasibility of implementing a CD approach for developing a safe and effective LASV live-attenuated vaccine candidate. Moreover, rLASV-GPC/CD might provide investigators with a tool to safely study LASV outside maximum (biosafety level 4) containment, which could accelerate the elucidation of basic aspects of the molecular and cell biology of LASV and the development of novel LASV medical countermeasures.IMPORTANCE Lassa virus (LASV) infects several hundred thousand people in Western Africa, resulting in many lethal Lassa fever (LF) cases. Licensed LF vaccines are not available, and anti-LF therapy is limited to off-label use of the nucleoside analog ribavirin with uncertain efficacy. We describe the generation of a novel live-attenuated LASV vaccine candidate. This vaccine candidate is based on mutating wild-type (WT) LASV in a key region of the viral genome, the glycoprotein precursor (GPC) gene. These mutations do not change the encoded GPC but interfere with its production in host cells. This mutated LASV (rLASV-GPC/CD) behaves like WT LASV (rLASV-WT) in cell culture, but in contrast to rLASV-WT, does not cause disease in inoculated guinea pigs. Guinea pigs immunized with rLASV-GPC/CD were protected against an otherwise lethal exposure to WT LASV. Our results support the testing of this candidate vaccine in nonhuman primate models of LF.