SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract

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 re...

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Bibliographic Details
Published in:Cell 2020-07, Vol.182 (2), p.429-446.e14
Main Authors: Hou, Yixuan J., Okuda, Kenichi, Edwards, Caitlin E., Martinez, David R., Asakura, Takanori, Dinnon, Kenneth H., Kato, Takafumi, Lee, Rhianna E., Yount, Boyd L., Mascenik, Teresa M., Chen, Gang, Olivier, Kenneth N., Ghio, Andrew, Tse, Longping V., Leist, Sarah R., Gralinski, Lisa E., Schäfer, Alexandra, Dang, Hong, Gilmore, Rodney, Nakano, Satoko, Sun, Ling, Fulcher, M. Leslie, Livraghi-Butrico, Alessandra, Nicely, Nathan I., Cameron, Mark, Cameron, Cheryl, Kelvin, David J., de Silva, Aravinda, Margolis, David M., Markmann, Alena, Bartelt, Luther, Zumwalt, Ross, Martinez, Fernando J., Salvatore, Steven P., Borczuk, Alain, Tata, Purushothama R., Sontake, Vishwaraj, Kimple, Adam, Jaspers, Ilona, O’Neal, Wanda K., Randell, Scott H., Boucher, Richard C., Baric, Ralph S.
Format: Article
Language:English
Subjects:
ACE2
Aged
Angiotensin-Converting Enzyme 2
Animals
Antibodies, Monoclonal - immunology
Antibodies, Neutralizing - immunology
Betacoronavirus - genetics
Betacoronavirus - immunology
Betacoronavirus - pathogenicity
Cell Line
Cells, Cultured
Chlorocebus aethiops
Coronavirus Infections - immunology
Coronavirus Infections - pathology
Coronavirus Infections - therapy
Coronavirus Infections - virology
COVID-19
COVID-19 Serotherapy
Cystic Fibrosis - pathology
DNA, Recombinant
Female
Furin - metabolism
Humans
Immunization, Passive
infectious clone
Lung - metabolism
Lung - pathology
Lung - virology
Male
Middle Aged
nasal infection
Nasal Mucosa - metabolism
Nasal Mucosa - pathology
Nasal Mucosa - virology
neutralization assay
Pandemics
Peptidyl-Dipeptidase A - metabolism
Pneumonia, Viral - immunology
Pneumonia, Viral - pathology
Pneumonia, Viral - virology
primary cells
reporter virus
Respiratory System - pathology
Respiratory System - virology
respiratory tropism
reverse genetics
Reverse Genetics - methods
SARS-CoV-2
Serine Endopeptidases - metabolism
Vero Cells
Virulence
Virus Replication
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Summary: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. [Display omitted] •A SARS-CoV-2 infectious cDNA clone and reporter viruses are generated•SARS-CoV-2 and SARS-CoV neutralization assays show limited cross neutralization•SARS-CoV-2 shows a gradient infectivity from the proximal to distal respiratory tract•Ciliated airway cells and AT-2 cells are primary targets for SARS-CoV-2 infection Hou et al. present a reverse genetics system for SARS-CoV-2, which is then used to make reporter viruses to quantify the ability of patient sera and antibodies to neutralize infectious virus and to examine viral tropism along the human respiratory tract.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2020.05.042