Synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice
Synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice
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DOI：
10.1128/jvi.00505-07
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发表时间：
2007-07-01
影响因子：
5.4
通讯作者：
Baric, Ralph
中科院分区：
文献类型：
--
作者：
Rockx, Barry;Sheahan, Timothy;Baric, Ralph
严重急性呼吸综合征(SARS)疫情的特点是老年人死亡率高。控制老年人群易感性增加的分子机制尚不清楚，需要强大的动物模型来概括随着年龄增加而增加的致病表型。利用合成生物学和反向遗传学，我们描述了一组携带变异的尖峰糖蛋白的SARS冠状病毒(SARS-CoV)毒株的构建，这些变异的尖峰糖蛋白代表在果子狸和浣熊身上发现的人畜共患病株，以及横跨SARS-CoV流行的早期、中期和晚期的分离株。重组病毒在细胞培养中有效复制，并对抗体中和表现出不同的敏感性。人而不是人畜共患变种在人的呼吸道上皮细胞培养中有效复制，支持了早期的假设，即人畜共患分离株对人类的致病性较低，但可以进化为高致病性菌株。所有病毒都有效复制，但没有一种病毒在幼年动物中产生临床疾病或死亡。相比之下，接种了果子狸HC/SZ/61/03株或人类早期GZ02变异株的12个月龄小鼠出现了严重的临床疾病、弥漫性肺泡损伤、肺透明膜形成、肺泡炎和死亡，但没有接种相关的中晚期流行株或浣熊犬株。这一组携带人畜共患病和人类流行尖峰糖蛋白的SARS-CoV重组体将提供异源挑战模型，用于测试针对人畜共患病重新引入的疫苗效力，并提供适当的模型系统，以阐明导致老年人更严重和致命的SARS-CoV疾病和急性呼吸窘迫的复杂病毒-宿主相互作用。
The severe acute respiratory syndrome (SARS) epidemic was characterized by high mortality rates in the elderly. The molecular mechanisms that govern enhanced susceptibility of elderly populations are not known, and robust animal models are needed that recapitulate the increased pathogenic phenotype noted with increasing age. Using synthetic biology and reverse genetics, we describe the construction of a panel of isogenic SARS coronavirus (SARS-CoV) strains bearing variant spike glycoproteins that are representative of zoonotic strains found in palm civets and raccoon dogs, as well as isolates spanning the early, middle, and late phases of the SARS-CoV epidemic. The recombinant viruses replicated efficiently in cell culture and demonstrated variable sensitivities to neutralization with antibodies. The human but not the zoonotic variants replicated efficiently in human airway epithelial cultures, supporting earlier hypotheses that zoonotic isolates are less pathogenic in humans but can evolve into highly pathogenic strains. All viruses replicated efficiently, but none produced clinical disease or death in young animals. In contrast, severe clinical disease, diffuse alveolar damage, hyaline membrane formation, alveolitis, and death were noted in 12-month-old mice inoculated with the palm civet HC/SZ/61/03 strain or early-human-phase GZ02 variants but not with related middle- and late-phase epidemic or raccoon dog strains. This panel of SARS-CoV recombinants bearing zoonotic and human epidemic spike glycoproteins will provide heterologous challenge models for testing vaccine efficacy against zoonotic reintroductions as well as provide the appropriate model system for elucidating the complex virus-host interactions that contribute to more-severe and fatal SARS-CoV disease and acute respiratory distress in the elderly.