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

被引:1045
作者
Hou, Yixuan J. [1 ]
Okuda, Kenichi [3 ]
Edwards, Caitlin E. [1 ]
Martinez, David R. [1 ]
Asakura, Takanori [3 ]
Dinnon, Kenneth H., III [2 ]
Kato, Takafumi [3 ]
Lee, Rhianna E. [3 ]
Yount, Boyd L. [1 ]
Mascenik, Teresa M. [3 ]
Chen, Gang [3 ]
Olivier, Kenneth N. [16 ]
Ghio, Andrew [17 ]
Tse, Longping, V [1 ]
Leist, Sarah R. [1 ]
Gralinski, Lisa E. [1 ]
Schafer, Alexandra [1 ]
Dang, Hong [3 ]
Gilmore, Rodney [3 ]
Nakano, Satoko [3 ]
Sun, Ling [3 ]
Fulcher, M. Leslie [3 ]
Livraghi-Butrico, Alessandra [3 ]
Nicely, Nathan, I [4 ]
Cameron, Mark [11 ]
Cameron, Cheryl [12 ]
Kelvin, David J. [10 ,18 ]
de Silva, Aravinda [2 ]
Margolis, David M. [2 ,5 ,6 ]
Markmann, Alena [5 ]
Bartelt, Luther [5 ]
Zumwalt, Ross [13 ]
Martinez, Fernando J. [14 ]
Salvatore, Steven P. [15 ]
Borczuk, Alain [15 ]
Tata, Purushothama R. [9 ]
Sontake, Vishwaraj [9 ]
Kimple, Adam [7 ]
Jaspers, Ilona [8 ]
O'Neal, Wanda K. [3 ]
Randell, Scott H. [3 ]
Boucher, Richard C. [3 ]
Baric, Ralph S. [1 ,2 ]
机构
[1] Univ N Carolina, Dept Epidemiol, Chapel Hill, NC 27515 USA
[2] Univ N Carolina, Dept Microbiol & Immunol, Chapel Hill, NC 27515 USA
[3] Univ N Carolina, Marsico Lung Inst, Chapel Hill, NC 27515 USA
[4] Univ N Carolina, Prot Express & Purificat Core, Chapel Hill, NC 27515 USA
[5] Univ N Carolina, Dept Med, Chapel Hill, NC 27515 USA
[6] Univ N Carolina, HIV Cure Ctr, Chapel Hill, NC 27515 USA
[7] Univ N Carolina, Dept Otolaryngol, Chapel Hill, NC 27515 USA
[8] Univ N Carolina, Dept Pediat, Chapel Hill, NC 27515 USA
[9] Duke Univ, Med Ctr, Dept Cell Biol, Regenerat Next Initiat, Durham, NC 27710 USA
[10] Dalhousie Univ, Canadian Ctr Vaccinol, Dept Microbiol & Immunol, Halifax, NS, Canada
[11] Case Western Reserve Univ, Dept Populat & Quantitat Hlth Sci, Cleveland, OH 44106 USA
[12] Case Western Reserve Univ, Dept Nutr, Cleveland, OH 44106 USA
[13] Univ New Mexico, Dept Pathol, Albuquerque, NM 87131 USA
[14] Cornell Univ, Joan & Sanford I Weill Med Coll, Div Pulm & Crit Care Med, New York, NY 10021 USA
[15] Cornell Univ, Joan & Sanford I Weill Med Coll, Dept Pathol, New York, NY 10021 USA
[16] NHLBI, Lab Chron Airway Infect, Pulm Branch, NIH, Bldg 10, Bethesda, MD 20892 USA
[17] US EPA, Natl Hlth & Environm Effects Res Lab, Chapel Hill, NC USA
[18] Shantou Univ, Lab Immunol, Med Coll, Shantou, Guangdong, Peoples R China
关键词
CORONAVIRUS SPIKE PROTEIN; ENZYME; 2; ACE2; SARS CORONAVIRUS; CELL ENTRY; CDNA-CLONE; MERS-COV; VIRUS; NEUTRALIZATION; ACTIVATION; ASPIRATION;
D O I
10.1016/j.cell.2020.05.042
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:429 / +
页数:32
相关论文
共 107 条
[1]  
Agnihothram S, 2018, J VIROL, V92, DOI [10.1128/JVI.00027-18, 10.1128/jvi.00027-18]
[2]   Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue [J].
Aguiar, Jennifer A. ;
Tremblay, Benjamin J-M ;
Mansfield, Michael J. ;
Woody, Owen ;
Lobb, Briallen ;
Banerjee, Arinjay ;
Chandiramohan, Abiram ;
Tiessen, Nicholas ;
Cao, Quynh ;
Dvorkin-Gheva, Anna ;
Revill, Spencer ;
Miller, Matthew S. ;
Carlsten, Christopher ;
Organ, Louise ;
Joseph, Chitra ;
John, Alison ;
Hanson, Paul ;
Austin, Richard C. ;
McManus, Bruce M. ;
Jenkins, Gisli ;
Mossman, Karen ;
Ask, Kjetil ;
Doxey, Andrew C. ;
Hirota, Jeremy A. .
EUROPEAN RESPIRATORY JOURNAL, 2020, 56 (03)
[3]   Engineering the largest RNA virus genome as an infectious bacterial artificial chromosome [J].
Almazán, F ;
González, JM ;
Pénzes, Z ;
Izeta, A ;
Calvo, E ;
Plana-Durán, J ;
Enjuanes, L .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (10) :5516-5521
[4]  
AMBERSON JB, 1954, B JOHNS HOPKINS HOSP, V94, P227
[5]   The proximal origin of SARS-CoV-2 [J].
Andersen, Kristian G. ;
Rambaut, Andrew ;
Lipkin, W. Ian ;
Holmes, Edward C. ;
Garry, Robert F. .
NATURE MEDICINE, 2020, 26 (04) :450-452
[6]  
[Anonymous], MMWR MORB MORTAL WKL, V69, P382
[7]   COVID-19 for the Cardiologist Basic Virology, Epidemiology, Cardiac Manifestations, and Potential Therapeutic Strategies [J].
Atri, Deepak ;
Siddiqi, Hasan K. ;
Lang, Joshua P. ;
Nauffal, Victor ;
Morrow, David A. ;
Bohula, Erin A. .
JACC-BASIC TO TRANSLATIONAL SCIENCE, 2020, 5 (05) :518-536
[8]   Fitting Linear Mixed-Effects Models Using lme4 [J].
Bates, Douglas ;
Maechler, Martin ;
Bolker, Benjamin M. ;
Walker, Steven C. .
JOURNAL OF STATISTICAL SOFTWARE, 2015, 67 (01) :1-48
[9]   Characterization of a Pathogenic Full-Length cDNA Clone and Transmission Model for Porcine Epidemic Diarrhea Virus Strain PC22A [J].
Beall, Anne ;
Yount, Boyd ;
Lin, Chun-Ming ;
Hou, Yixuan ;
Wang, Qiuhong ;
Saif, Linda ;
Baric, Ralph .
MBIO, 2016, 7 (01)
[10]   Detection of airborne severe acute respiratory syndrome (SARS) coronavirus and environmental contamination in SARS outbreak units [J].
Booth, TF ;
Kournikakis, B ;
Bastien, N ;
Ho, J ;
Kobasa, D ;
Stadnyk, L ;
Li, Y ;
Spence, M ;
Paton, S ;
Henry, B ;
Mederski, B ;
White, D ;
Low, DE ;
McGeer, A ;
Simor, A ;
Vearncombe, M ;
Downey, J ;
Jamieson, FB ;
Tang, P ;
Plummer, F .
JOURNAL OF INFECTIOUS DISEASES, 2005, 191 (09) :1472-1477