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Cell
. 2020 May 27;S0092-8674(20)30675-9.
doi: 10.1016/j.cell.2020.05.042. Online ahead of print.
SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract
Yixuan J Hou 1 , Kenichi Okuda 2 , Caitlin E Edwards 1 , David R Martinez 1 , Takanori Asakura 2 , Kenneth H Dinnon 3rd 3 , Takafumi Kato 2 , Rhianna E Lee 2 , Boyd L Yount 1 , Teresa M Mascenik 2 , Gang Chen 2 , Kenneth N Olivier 4 , Andrew Ghio 5 , Longping V Tse 1 , Sarah R Leist 1 , Lisa E Gralinski 1 , Alexandra Sch?fer 1 , Hong Dang 2 , Rodney Gilmore 2 , Satoko Nakano 2 , Ling Sun 2 , M Leslie Fulcher 2 , Alessandra Livraghi-Butrico 2 , Nathan I Nicely 6 , Mark Cameron 7 , Cheryl Cameron 8 , David J Kelvin 9 , Aravinda de Silva 3 , David M Margolis 10 , Alena Markmann 11 , Luther Bartelt 11 , Ross Zumwalt 12 , Fernando J Martinez 13 , Steven P Salvatore 14 , Alain Borczuk 14 , Purushothama R Tata 15 , Vishwaraj Sontake 15 , Adam Kimple 16 , Ilona Jaspers 17 , Wanda K O'Neal 2 , Scott H Randell 2 , Richard C Boucher 18 , Ralph S Baric 19
Affiliations
- PMID: 32526206
- DOI: 10.1016/j.cell.2020.05.042
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.
Keywords: ACE2; COVID-19; SARS-CoV-2; infectious clone; nasal infection; neutralization assay; primary cells; reporter virus; respiratory tropism; reverse genetics.