Molecular Engineering of a Mammarenavirus with Unbreachable Attenuation.
Molecular Engineering of a Mammarenavirus with Unbreachable Attenuation.
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具有不可破坏的减毒能力的乳腺病毒的分子工程。
DOI：
10.1128/jvi.01385-22
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发表时间：
2023
影响因子：
5.4
通讯作者：
delaTorre,JuanC
中科院分区：
文献类型：
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作者：
Sakabe,Saori;Cubitt,Beatrice;Martinez-Sobrido,Luis;delaTorre,JuanC
几种乳房肾病毒在人类中引起严重的出血热（HF）疾病，并在其流行地区造成重要的公共卫生问题。没有美国（US）食品和药物管理局（FDA）批准的乳房炎病毒疫苗，目前的抗乳房炎病毒治疗仅限于利巴韦林的标签外使用，其疗效有限。哺乳动物肾病毒是具有双节段负链RNA基因组的包膜病毒。每个基因组片段包含两个开放阅读框（ORF），由非编码基因间区（IGR）分隔。大（L）片段编码RNA依赖性RNA聚合酶、L蛋白和Z基质蛋白，而小（S）片段编码表面糖蛋白前体（GPC）和核蛋白（NP）。在本研究中，我们记录的原型mammarenavirus淋巴细胞性脉络丛脑膜炎病毒（LCMV）表达密码子去优化（CD）GPC和包含IGR的S段在S和L段（rLCMV/IGR-CD）的重组形式的生成。我们表明，rLCMV/IGR-CD在C57 BL/6（B6）小鼠中完全减毒，但能够在单次给药后提供完全保护，以对抗LCMV的致死性攻击。重要的是，rLCMV/IGR-CD表现出不可破坏的减毒，其安全的实施作为减毒活疫苗（LAV）。重要几个mammarenaviruses引起严重的疾病，在人类和造成重要的公共卫生问题，在其地区的地方性。目前，没有FDA许可的乳房炎病毒疫苗可用，抗乳房炎病毒治疗仅限于利巴韦林的标签外使用，其疗效存在争议。在这里，我们描述的原型mammarenavirus淋巴细胞性脉络丛脑膜炎病毒（rLCMV）的重组版本的生成相结合的密码子去优化（CD）GPC和非编码基因间区（IGR）的S段在S和L基因组片段，称为rLCMV/IGR-CD的功能。我们提出的证据表明，rLCMV/IGR-CD具有优良的安全性和保护效力的特点，减毒活疫苗（LAV）。重要的是，rLCMV/IGR-CD在共感染小鼠中防止产生具有增加的毒力的LCMV抑制剂。我们的研究结果证明了一种明确的分子策略，用于产生能够在单次免疫后引发长期保护性免疫的乳腺炎病毒LAV候选物，同时表现出独特的增强安全性特征，包括不可破坏的减毒。
Several mammarenaviruses cause severe hemorrhagic fever (HF) disease in humans and pose important public health problems in their regions of endemicity. There are no United States (US) Food and Drug Administration (FDA)-approved mammarenavirus vaccines, and current anti-mammarenavirus therapy is limited to an off-label use of ribavirin that has limited efficacy. Mammarenaviruses are enveloped viruses with a bi-segmented negative-strand RNA genome. Each genome segment contains two open reading frames (ORF) separated by a noncoding intergenic region (IGR). The large (L) segment encodes the RNA dependent RNA polymerase, L protein, and the Z matrix protein, whereas the small (S) segment encodes the surface glycoprotein precursor (GPC) and nucleoprotein (NP). In the present study, we document the generation of a recombinant form of the prototypic mammarenavirus lymphocytic choriomeningitis virus (LCMV) expressing a codon deoptimized (CD) GPC and containing the IGR of the S segment in both the S and L segments (rLCMV/IGR-CD). We show that rLCMV/IGR-CD is fully attenuated in C57BL/6 (B6) mice but able to provide complete protection upon a single administration against a lethal challenge with LCMV. Importantly, rLCMV/IGR-CD exhibited an unbreachable attenuation for its safe implementation as a live-attenuated vaccine (LAV).IMPORTANCESeveral mammarenaviruses cause severe disease in humans and pose important public health problems in their regions of endemicity. Currently, no FDA-licensed mammarenavirus vaccines are available, and anti-mammarenaviral therapy is limited to an off-label use of ribavirin whose efficacy is controversial. Here, we describe the generation of recombinant version of the prototypic mammarenavirus lymphocytic choriomeningitis virus (rLCMV) combining the features of a codon deoptimized (CD) GPC and the noncoding intergenic region (IGR) of the S segment in both S and L genome segments, called rLCMV/IGR-CD. We present evidence that rLCMV/IGR-CD has excellent safety and protective efficacy features as live-attenuated vaccine (LAV). Importantly, rLCMV/IGR-CD prevents, in coinfected mice, the generation of LCMV reassortants with increased virulence. Our findings document a well-defined molecular strategy for the generation of mammarenavirus LAV candidates able to trigger long-term protective immunity, upon a single immunization, while exhibiting unique enhanced safety features, including unbreachable attenuation.