SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract
SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract
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DOI：
10.1016/j.cell.2020.05.042
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发表时间：
2020-07-23
期刊：
影响因子：
64.5
通讯作者：
Baric, Ralph S.
中科院分区：
文献类型：
--
作者：
Hou, Yixuan J.;Okuda, Kenichi;Baric, Ralph S.
2019年冠状病毒病(新冠肺炎)临床谱可变的获取方式和原因尚不清楚。我们利用反向遗传学系统生成绿色荧光蛋白报告病毒来研究严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的致病机理，并利用荧光素酶报告病毒来证明从SARS和新冠肺炎患者收集的血清表现出有限的交叉冠状病毒中和。高灵敏度RNA原位图谱显示血管紧张素转换酶2(ACE2)在鼻部表达最高，下呼吸道表达减少，与SARS-CoV-2在近端(高)和远端(低)肺上皮细胞培养中显著的感染梯度平行。新冠肺炎尸检发现了局限性病变，与培养数据一致，分别在呼吸道和肺泡区发现了感染SARS-CoV-2的纤毛细胞和2型肺泡细胞。这些发现突出了鼻腔对SARS-CoV-2的易感性，在SARS-CoV-2的发病机制中，可能随后通过吸入介导的病毒播撒到肺部。这些试剂为研究病毒与宿主在保护性免疫、宿主易感性和病毒致病机制中的相互作用奠定了基础。
The mode of acquisition and causes for the variable clinical spectrum of coronavirus disease 2019 (COVID-19) remain unknown. We utilized a reverse genetics system to generate a GFP reporter virus to explore severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pathogenesis and a luciferase reporter virus to demonstrate sera collected from SARS and COVID-19 patients exhibited limited cross-CoV neutralization. High-sensitivity RNA in situ mapping revealed the highest angiotensin-converting enzyme 2 (ACE2) expression in the nose with decreasing expression throughout the lower respiratory tract, paralleled by a striking gradient of SARS-CoV-2 infection in proximal (high) versus distal (low) pulmonary epithelial cultures. COVID-19 autopsied lung studies identified focal disease and, congruent with culture data, SARS-CoV-2-infected ciliated and type 2 pneumocyte cells in airway and alveolar regions, respectively. These findings highlight the nasal susceptibility to SARS-CoV-2 with likely subsequent aspiration-mediated virus seeding to the lung in SARS-CoV-2 pathogenesis. These reagents provide a foundation for investigations into virus-host interactions in protective immunity, host susceptibility, and virus pathogenesis.