SARS-CoV-2 variant Alpha has a spike-dependent replication advantage over the ancestral B.1 strain in human cells with low ACE2 expression.

Autor: Niemeyer D; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany., Stenzel S; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Veith T; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany., Schroeder S; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Friedmann K; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Weege F; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Trimpert J; Institut für Virologie, Freie Universität Berlin, Berlin, Germany., Heinze J; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany., Richter A; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Jansen J; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Emanuel J; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Kazmierski J; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Pott F; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Jeworowski LM; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Olmer R; Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH - Center for Translational Regenerative Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover Medical School, Hannover, Germany., Jaboreck MC; Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH - Center for Translational Regenerative Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover Medical School, Hannover, Germany., Tenner B; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Papies J; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Walper F; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Schmidt ML; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Heinemann N; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Möncke-Buchner E; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Baumgardt M; Department of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany., Hoffmann K; Department of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany., Widera M; Institute for Medical Virology, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany., Thao TTN; Institute of Virology and Immunology, Bern, Switzerland., Balázs A; Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany., Schulze J; Unit 17 'Influenza and other Respiratory Viruses', Robert Koch Institute, Berlin, Germany., Mache C; Unit 17 'Influenza and other Respiratory Viruses', Robert Koch Institute, Berlin, Germany., Jones TC; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany., Morkel M; Institute of Pathology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt - Universität zu Berlin, Berlin, Germany.; BIH Bioportal Single Cells, Berlin Institute of Health at Charité Universitätsmedizin Berlin, Berlin, Germany., Ciesek S; Institute for Medical Virology, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany.; German Center for Infection Research, DZIF, Braunschweig, Germany.; Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Branch Translational Medicine and Pharmacology, Frankfurt am Main, Germany., Hanitsch LG; Institute of Medical Immunology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany., Mall MA; Institut für Virologie, Freie Universität Berlin, Berlin, Germany.; Department of Pediatric Respiratory Medicine, Immunology and Critical Care Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Centre for Lung Research (DZL), associated partner Charité, Berlin, Germany., Hocke AC; Department of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany., Thiel V; Institute of Virology and Immunology, Bern, Switzerland., Osterrieder K; Berlin Institute of Health, Berlin, Germany.; Department of Infectious Diseases and Public Health, Jockey Club College of Veterinary Medicine and Life Sciences, City University of Hong Kong, Kowloon Tong, Hong Kong., Wolff T; Unit 17 'Influenza and other Respiratory Viruses', Robert Koch Institute, Berlin, Germany., Martin U; Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH - Center for Translational Regenerative Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover Medical School, Hannover, Germany., Corman VM; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany.; Labor Berlin - Charité Vivantes GmbH, Berlin, Germany., Müller MA; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany., Goffinet C; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Drosten C; Institute of Virology, Campus Charité Mitte, Charité - Universitätsmedizin Berlin, Berlin, Germany.; German Center for Infection Research, associated partner Charité, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany.; Labor Berlin - Charité Vivantes GmbH, Berlin, Germany.
Jazyk: angličtina
Zdroj: PLoS biology [PLoS Biol] 2022 Nov 16; Vol. 20 (11), pp. e3001871. Date of Electronic Publication: 2022 Nov 16 (Print Publication: 2022).
DOI: 10.1371/journal.pbio.3001871
Abstrakt: Epidemiological data demonstrate that Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) Alpha and Delta are more transmissible, infectious, and pathogenic than previous variants. Phenotypic properties of VOC remain understudied. Here, we provide an extensive functional study of VOC Alpha replication and cell entry phenotypes assisted by reverse genetics, mutational mapping of spike in lentiviral pseudotypes, viral and cellular gene expression studies, and infectivity stability assays in an enhanced range of cell and epithelial culture models. In almost all models, VOC Alpha spread less or equally efficiently as ancestral (B.1) SARS-CoV-2. B.1. and VOC Alpha shared similar susceptibility to serum neutralization. Despite increased relative abundance of specific sgRNAs in the context of VOC Alpha infection, immune gene expression in infected cells did not differ between VOC Alpha and B.1. However, inferior spreading and entry efficiencies of VOC Alpha corresponded to lower abundance of proteolytically cleaved spike products presumably linked to the T716I mutation. In addition, we identified a bronchial cell line, NCI-H1299, which supported 24-fold increased growth of VOC Alpha and is to our knowledge the only cell line to recapitulate the fitness advantage of VOC Alpha compared to B.1. Interestingly, also VOC Delta showed a strong (595-fold) fitness advantage over B.1 in these cells. Comparative analysis of chimeric viruses expressing VOC Alpha spike in the backbone of B.1, and vice versa, showed that the specific replication phenotype of VOC Alpha in NCI-H1299 cells is largely determined by its spike protein. Despite undetectable ACE2 protein expression in NCI-H1299 cells, CRISPR/Cas9 knock-out and antibody-mediated blocking experiments revealed that multicycle spread of B.1 and VOC Alpha required ACE2 expression. Interestingly, entry of VOC Alpha, as opposed to B.1 virions, was largely unaffected by treatment with exogenous trypsin or saliva prior to infection, suggesting enhanced resistance of VOC Alpha spike to premature proteolytic cleavage in the extracellular environment of the human respiratory tract. This property may result in delayed degradation of VOC Alpha particle infectivity in conditions typical of mucosal fluids of the upper respiratory tract that may be recapitulated in NCI-H1299 cells closer than in highly ACE2-expressing cell lines and models. Our study highlights the importance of cell model evaluation and comparison for in-depth characterization of virus variant-specific phenotypes and uncovers a fine-tuned interrelationship between VOC Alpha- and host cell-specific determinants that may underlie the increased and prolonged virus shedding detected in patients infected with VOC Alpha.
Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Technische Universität Berlin, Freie Universität Berlin and Charité - Universitätsmedizin have filed a patent application for siRNAs inhibiting SARS-CoV-2 replication with DN as co-author. MAMü and VMC are named together with Charité - Universitätsmedizin Berlin and Euroimmun Medizinische Labordiagnostika AG on a patent application (EP3715847) filed recently regarding the diagnostic of SARS-CoV-2 by antibody testing. The other authors declare no competing interests.
(Copyright: © 2022 Niemeyer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje