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

By Yixuan J. Hou, K. Okuda, C. Edwards, David R. Martinez, T. Asakura, K. Dinnon, T. Kato, Rhianna E. Lee, B. Yount, T. Mascenik and 33 more. 2020

1,406 citations recorded by the source corpus.

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.

Subfields: Epidemiology, Infectious Diseases

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In Epidemiology

1. Yoshihiro Kawaoka
2. Ralph S. Baric

In Infectious Diseases

1. Ralph S. Baric
2. Rachel L. Graham

Citations in the corpus

• Tracking Changes in SARS-CoV-2 Spike: Evidence that D614G Increases Infectivity of the COVID-19 Virus (2020, neutral): “We encourage others to study this phenomenon in greater detail with a wild-type virus in natural infection and varied target cells (Hou et al., 2020) and in relevant animal models.”
• SARS-CoV-2 D614G Variant Exhibits Efficient Replication ex vivo and Transmission in vivo (2020, methodology): “epithelial (LAE)] and bronchiolar [small airway epithelial (SAE)] cells were isolated from freshly excised normal human tissues obtained from transplant donors under UNC Institutional Review Board (IRB)approved protocol (#03-1396) and cultured in air-liquid interface (ALI) media, as previously descr”
• A mouse-adapted model of SARS-CoV-2 to test COVID-19 countermeasures (2020, methodology): “Using molecular modelling and reverse genetics 22 , we altered the SARS-CoV-2 RBD to enable viral entry via mouse ACE2, highlighting the precision of the structure-based predictions.”
• SARS-CoV-2 mRNA Vaccine Design Enabled by Prototype Pathogen Preparedness (2020, neutral): “SARS-CoV-2 MA contains the substitutions Q498Y/P499T in the receptor-binding domain 31 .”