tetano
Editor, Senior Moderator
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[SUP] 1 [/SUP], Kenichi Okuda[SUP] 2 [/SUP], Caitlin E Edwards[SUP] 1 [/SUP], David R Martinez[SUP] 1 [/SUP], Takanori Asakura[SUP] 2 [/SUP], Kenneth H Dinnon 3rd[SUP] 3 [/SUP], Takafumi Kato[SUP] 2 [/SUP], Rhianna E Lee[SUP] 2 [/SUP], Boyd L Yount[SUP] 1 [/SUP], Teresa M Mascenik[SUP] 2 [/SUP], Gang Chen[SUP] 2 [/SUP], Kenneth N Olivier[SUP] 4 [/SUP], Andrew Ghio[SUP] 5 [/SUP], Longping V Tse[SUP] 1 [/SUP], Sarah R Leist[SUP] 1 [/SUP], Lisa E Gralinski[SUP] 1 [/SUP], Alexandra Sch?fer[SUP] 1 [/SUP], Hong Dang[SUP] 2 [/SUP], Rodney Gilmore[SUP] 2 [/SUP], Satoko Nakano[SUP] 2 [/SUP], Ling Sun[SUP] 2 [/SUP], M Leslie Fulcher[SUP] 2 [/SUP], Alessandra Livraghi-Butrico[SUP] 2 [/SUP], Nathan I Nicely[SUP] 6 [/SUP], Mark Cameron[SUP] 7 [/SUP], Cheryl Cameron[SUP] 8 [/SUP], David J Kelvin[SUP] 9 [/SUP], Aravinda de Silva[SUP] 3 [/SUP], David M Margolis[SUP] 10 [/SUP], Alena Markmann[SUP] 11 [/SUP], Luther Bartelt[SUP] 11 [/SUP], Ross Zumwalt[SUP] 12 [/SUP], Fernando J Martinez[SUP] 13 [/SUP], Steven P Salvatore[SUP] 14 [/SUP], Alain Borczuk[SUP] 14 [/SUP], Purushothama R Tata[SUP] 15 [/SUP], Vishwaraj Sontake[SUP] 15 [/SUP], Adam Kimple[SUP] 16 [/SUP], Ilona Jaspers[SUP] 17 [/SUP], Wanda K O'Neal[SUP] 2 [/SUP], Scott H Randell[SUP] 2 [/SUP], Richard C Boucher[SUP] 18 [/SUP], Ralph S Baric[SUP] 19 [/SUP]
Affiliations
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.
. 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[SUP] 1 [/SUP], Kenichi Okuda[SUP] 2 [/SUP], Caitlin E Edwards[SUP] 1 [/SUP], David R Martinez[SUP] 1 [/SUP], Takanori Asakura[SUP] 2 [/SUP], Kenneth H Dinnon 3rd[SUP] 3 [/SUP], Takafumi Kato[SUP] 2 [/SUP], Rhianna E Lee[SUP] 2 [/SUP], Boyd L Yount[SUP] 1 [/SUP], Teresa M Mascenik[SUP] 2 [/SUP], Gang Chen[SUP] 2 [/SUP], Kenneth N Olivier[SUP] 4 [/SUP], Andrew Ghio[SUP] 5 [/SUP], Longping V Tse[SUP] 1 [/SUP], Sarah R Leist[SUP] 1 [/SUP], Lisa E Gralinski[SUP] 1 [/SUP], Alexandra Sch?fer[SUP] 1 [/SUP], Hong Dang[SUP] 2 [/SUP], Rodney Gilmore[SUP] 2 [/SUP], Satoko Nakano[SUP] 2 [/SUP], Ling Sun[SUP] 2 [/SUP], M Leslie Fulcher[SUP] 2 [/SUP], Alessandra Livraghi-Butrico[SUP] 2 [/SUP], Nathan I Nicely[SUP] 6 [/SUP], Mark Cameron[SUP] 7 [/SUP], Cheryl Cameron[SUP] 8 [/SUP], David J Kelvin[SUP] 9 [/SUP], Aravinda de Silva[SUP] 3 [/SUP], David M Margolis[SUP] 10 [/SUP], Alena Markmann[SUP] 11 [/SUP], Luther Bartelt[SUP] 11 [/SUP], Ross Zumwalt[SUP] 12 [/SUP], Fernando J Martinez[SUP] 13 [/SUP], Steven P Salvatore[SUP] 14 [/SUP], Alain Borczuk[SUP] 14 [/SUP], Purushothama R Tata[SUP] 15 [/SUP], Vishwaraj Sontake[SUP] 15 [/SUP], Adam Kimple[SUP] 16 [/SUP], Ilona Jaspers[SUP] 17 [/SUP], Wanda K O'Neal[SUP] 2 [/SUP], Scott H Randell[SUP] 2 [/SUP], Richard C Boucher[SUP] 18 [/SUP], Ralph S Baric[SUP] 19 [/SUP]
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.