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

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

doi:10.1016/j.cell.2020.05.042

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

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 NakanoLing 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

Medicine

Research output: Contribution to journal › Article › peer-review

1101 Citations (Scopus)

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
Pages (from-to)	429-446.e14
Journal	Cell
Volume	182
Issue number	2
DOIs	https://doi.org/10.1016/j.cell.2020.05.042
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

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10.1016/j.cell.2020.05.042

Cite this

Hou, Y. J., Okuda, K., Edwards, C. E., Martinez, D. R., Asakura, T., Dinnon, K. H., Kato, T., Lee, R. E., Yount, B. L., Mascenik, T. M., Chen, G., Olivier, K. N., Ghio, A., Tse, L. V., Leist, S. R., Gralinski, L. E., Schäfer, A., Dang, H., Gilmore, R., ... Baric, R. S. (2020). SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract. Cell, 182(2), 429-446.e14. https://doi.org/10.1016/j.cell.2020.05.042

Hou, YJ, Okuda, K, Edwards, CE, Martinez, DR, Asakura, T, Dinnon, KH, Kato, T, Lee, RE, Yount, BL, Mascenik, TM, Chen, G, Olivier, KN, Ghio, A, Tse, LV, Leist, SR, Gralinski, LE, Schäfer, A, Dang, H, Gilmore, R, Nakano, S, Sun, L, Fulcher, ML, Livraghi-Butrico, A, Nicely, NI, Cameron, M, Cameron, C, Kelvin, DJ, de Silva, A, Margolis, DM, Markmann, A, Bartelt, L, Zumwalt, R, Martinez, FJ, Salvatore, SP, Borczuk, A, Tata, PR, Sontake, V, Kimple, A, Jaspers, I, O'Neal, WK, Randell, SH, Boucher, RC & Baric, RS 2020, 'SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract', Cell, vol. 182, no. 2, pp. 429-446.e14. https://doi.org/10.1016/j.cell.2020.05.042

@article{bad573cfe2e341628b82b972510993db,

title = "SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract",

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.",

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

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: {\textcopyright} 2020 Elsevier Inc.",

year = "2020",

month = jul,

day = "23",

doi = "10.1016/j.cell.2020.05.042",

language = "English",

volume = "182",

pages = "429--446.e14",

journal = "Cell",

issn = "0092-8674",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract

AU - Hou, Yixuan J.

AU - Okuda, Kenichi

AU - Edwards, Caitlin E.

AU - Martinez, David R.

AU - Asakura, Takanori

AU - Dinnon, Kenneth H.

AU - Kato, Takafumi

AU - Lee, Rhianna E.

AU - Yount, Boyd L.

AU - Mascenik, Teresa M.

AU - Chen, Gang

AU - Olivier, Kenneth N.

AU - Ghio, Andrew

AU - Tse, Longping V.

AU - Leist, Sarah R.

AU - Gralinski, Lisa E.

AU - Schäfer, Alexandra

AU - Dang, Hong

AU - Gilmore, Rodney

AU - Nakano, Satoko

AU - Sun, Ling

AU - Fulcher, M. Leslie

AU - Livraghi-Butrico, Alessandra

AU - Nicely, Nathan I.

AU - Cameron, Mark

AU - Cameron, Cheryl

AU - Kelvin, David J.

AU - de Silva, Aravinda

AU - Margolis, David M.

AU - Markmann, Alena

AU - Bartelt, Luther

AU - Zumwalt, Ross

AU - Martinez, Fernando J.

AU - Salvatore, Steven P.

AU - Borczuk, Alain

AU - Tata, Purushothama R.

AU - Sontake, Vishwaraj

AU - Kimple, Adam

AU - Jaspers, Ilona

AU - O'Neal, Wanda K.

AU - Randell, Scott H.

AU - Boucher, Richard C.

AU - Baric, Ralph S.

N1 - 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.

PY - 2020/7/23

Y1 - 2020/7/23

N2 - 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.

AB - 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.

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UR - http://www.scopus.com/inward/citedby.url?scp=85086399821&partnerID=8YFLogxK

U2 - 10.1016/j.cell.2020.05.042

DO - 10.1016/j.cell.2020.05.042

M3 - Article

C2 - 32526206

AN - SCOPUS:85086399821

SN - 0092-8674

VL - 182

SP - 429-446.e14

JO - Cell

JF - Cell

IS - 2

ER -

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

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