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

Cell (2020-06-12)
Yixuan J Hou, Kenichi Okuda, Caitlin E Edwards, David R Martinez, Takanori Asakura, Kenneth H Dinnon, Takafumi Kato, Rhianna E Lee, Boyd L Yount, Teresa M Mascenik, Gang Chen, Kenneth N Olivier, Andrew Ghio, Longping V Tse, Sarah R Leist, Lisa E Gralinski, Alexandra Schäfer, Hong Dang, Rodney Gilmore, Satoko Nakano, Ling Sun, M Leslie Fulcher, Alessandra Livraghi-Butrico, Nathan I Nicely, Mark Cameron, Cheryl Cameron, David J Kelvin, Aravinda de Silva, David M Margolis, Alena Markmann, Luther Bartelt, Ross Zumwalt, Fernando J Martinez, Steven P Salvatore, Alain Borczuk, Purushothama R Tata, Vishwaraj Sontake, Adam Kimple, Ilona Jaspers, Wanda K O'Neal, Scott H Randell, Richard C Boucher, Ralph S Baric
ABSTRACT

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.

MATERIALS
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