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.
Original language | English |
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Pages (from-to) | 429-446.e14 |
Journal | Cell |
Volume | 182 |
Issue number | 2 |
DOIs | |
Publication status | Published - Jul 23 2020 |
Bibliographical note
Funding Information:We would like to acknowledge funding sources from the National Allergy and Infectious Disease (NIAID), National Institution of Health (NIH) ( U19-AI100625 , R01-AI089728 , and U01-AI14964 ) to R.S.B. and the National Heart, Lung, and Blood Institute (NHLBI), NIH ( UH3-HL123645 , P01-HL110873 , R01-HL136961 , P30-DK065988-13 , and P01-HL108808 ) to R.C.B. K.O. is funded by the Cystic Fibrosis Foundation ( OKUDA10I0 ) and a research grant from Cystic Fibrosis Research Incorporation . D.R.M. is funded by NIH NIAID T32 AI007151 and a Burroughs Wellcome Fund Postdoctoral Enrichment Program Award . T.K. is funded by a Senior Research Training Fellowship ( RT-57362 ) of American Lung Association . A.J.K. is support by the National Center for Advancing Translational Sciences , NIH, through grant KL2TR002490 . P.R.T. received a Pathways to Independence award from the NHLBI / NIH ( R00HL127181 and R01HL146557 ) that partially supported this study. V.S. is supported by a postdoc fellowship from Regeneration Next Initiative at Duke University. We thank N.J. Thornburg at the CDC for providing us the SARS-CoV-2 clinical isolate WA1 strain. We are grateful for the technical support of Y. Escobar for HNE cultures and Lisa Morton for qPCR assays and to S. Weiss, Y. Park, J. Kuruc, and the UNC Blood Donor Center for COVID-19 serum sample preparation. The UNC Animal Histopathology & Laboratory Medicine Core is supported in part by an NCI Center Core Support Grant ( 5P30CA016086-41 ) to the UNC Lineberger Comprehensive Cancer Center. We thank E.C. Roe for assisting manuscript editing. Finally, we are grateful for the donors of primary cells and sera who made this study possible.
Funding Information:
We would like to acknowledge funding sources from the National Allergy and Infectious Disease (NIAID), National Institution of Health (NIH) (U19-AI100625, R01-AI089728, and U01-AI14964) to R.S.B. and the National Heart, Lung, and Blood Institute (NHLBI), NIH (UH3-HL123645, P01-HL110873, R01-HL136961, P30-DK065988-13, and P01-HL108808) to R.C.B. K.O. is funded by the Cystic Fibrosis Foundation (OKUDA10I0) and a research grant from Cystic Fibrosis Research Incorporation. D.R.M. is funded by NIH NIAID T32 AI007151 and a Burroughs Wellcome Fund Postdoctoral Enrichment Program Award. T.K. is funded by a Senior Research Training Fellowship (RT-57362) of American Lung Association. A.J.K. is support by the National Center for Advancing Translational Sciences, NIH, through grant KL2TR002490. P.R.T. received a Pathways to Independence award from the NHLBI/NIH (R00HL127181 and R01HL146557) that partially supported this study. V.S. is supported by a postdoc fellowship from Regeneration Next Initiative at Duke University. We thank N.J. Thornburg at the CDC for providing us the SARS-CoV-2 clinical isolate WA1 strain. We are grateful for the technical support of Y. Escobar for HNE cultures and Lisa Morton for qPCR assays and to S. Weiss, Y. Park, J. Kuruc, and the UNC Blood Donor Center for COVID-19 serum sample preparation. The UNC Animal Histopathology & Laboratory Medicine Core is supported in part by an NCI Center Core Support Grant (5P30CA016086-41) to the UNC Lineberger Comprehensive Cancer Center. We thank E.C. Roe for assisting manuscript editing. Finally, we are grateful for the donors of primary cells and sera who made this study possible. Conceptualization R.C.B. R.S.B, and S.H.R.; Investigation: Y.J.H. K.O. C.E.E. D.R.M. T.A. K.D.3, T.K. R.L. B.L.Y. T.M.M. G.C. K.N.O. A.G. L.V.T. S.R.L. L.E.G. A.S. H.D. R.G.S.N. L.S. L.F. W.K.O. and S.H.R.; Contribution to research materials: A.L.B. N.I.N. M.C. C.C. D.J.K. A.D.S. D.M.M. A.M. L.B. R.Z. F.J.M. S.P.S. A.B. P.R.T. V.S. A.K. I.J. and S.H.R. Writing ? original draft preparation: Y.J.H.; Writing ? review and editing: R.C.B. R.S.B. S.H.R. and W.K.O.; Visualization: Y.J.H. K.O. C.E.E. D.R.M. T.A. and T.K.; Funding acquisition: R.C.B. and R.S.B. The authors declare no competing financial interests.
Publisher Copyright:
© 2020 Elsevier Inc.
ASJC Scopus Subject Areas
- General Biochemistry,Genetics and Molecular Biology