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Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike

SARS-CoV-2 pathogenesis, vaccine, and therapeutic studies rely on the use of animals challenged with highly pathogenic virus stocks produced in cell cultures. Ideally, these virus stocks should be genetically and functionally similar to the original clinical isolate, retaining wild-type properties t...

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Autores principales: Baczenas, John James, Andersen, Hanne, Rashid, Sujatha, Yarmosh, David, Puthuveetil, Nikhita, Parker, Michael, Bradford, Rebecca, Florence, Clint, Stemple, Kimberly J., Lewis, Mark G., O’Connor, Shelby L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8704555/
https://www.ncbi.nlm.nih.gov/pubmed/34960703
http://dx.doi.org/10.3390/v13122434
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author Baczenas, John James
Andersen, Hanne
Rashid, Sujatha
Yarmosh, David
Puthuveetil, Nikhita
Parker, Michael
Bradford, Rebecca
Florence, Clint
Stemple, Kimberly J.
Lewis, Mark G.
O’Connor, Shelby L.
author_facet Baczenas, John James
Andersen, Hanne
Rashid, Sujatha
Yarmosh, David
Puthuveetil, Nikhita
Parker, Michael
Bradford, Rebecca
Florence, Clint
Stemple, Kimberly J.
Lewis, Mark G.
O’Connor, Shelby L.
author_sort Baczenas, John James
collection PubMed
description SARS-CoV-2 pathogenesis, vaccine, and therapeutic studies rely on the use of animals challenged with highly pathogenic virus stocks produced in cell cultures. Ideally, these virus stocks should be genetically and functionally similar to the original clinical isolate, retaining wild-type properties to be reliably used in animal model studies. It is well-established that SARS-CoV-2 isolates serially passaged on Vero cell lines accumulate mutations and deletions in the furin cleavage site; however, these can be eliminated when passaged on Calu-3 lung epithelial cell lines, as presented in this study. As numerous stocks of SARS-CoV-2 variants of concern are being grown in cell cultures with the intent for use in animal models, it is essential that propagation methods generate virus stocks that are pathogenic in vivo. Here, we found that the propagation of a B.1.351 SARS-CoV-2 stock on Calu-3 cells eliminated viruses that previously accumulated mutations in the furin cleavage site. Notably, there were alternative variants that accumulated at the same nucleotide positions in virus populations grown on Calu-3 cells at multiple independent facilities. When a Calu-3-derived B.1.351 virus stock was used to infect hamsters, the virus remained pathogenic and the Calu-3-specific variants persisted in the population. These results suggest that Calu-3-derived virus stocks are pathogenic but care should still be taken to evaluate virus stocks for newly arising mutations during propagation.
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spelling pubmed-87045552021-12-25 Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike Baczenas, John James Andersen, Hanne Rashid, Sujatha Yarmosh, David Puthuveetil, Nikhita Parker, Michael Bradford, Rebecca Florence, Clint Stemple, Kimberly J. Lewis, Mark G. O’Connor, Shelby L. Viruses Article SARS-CoV-2 pathogenesis, vaccine, and therapeutic studies rely on the use of animals challenged with highly pathogenic virus stocks produced in cell cultures. Ideally, these virus stocks should be genetically and functionally similar to the original clinical isolate, retaining wild-type properties to be reliably used in animal model studies. It is well-established that SARS-CoV-2 isolates serially passaged on Vero cell lines accumulate mutations and deletions in the furin cleavage site; however, these can be eliminated when passaged on Calu-3 lung epithelial cell lines, as presented in this study. As numerous stocks of SARS-CoV-2 variants of concern are being grown in cell cultures with the intent for use in animal models, it is essential that propagation methods generate virus stocks that are pathogenic in vivo. Here, we found that the propagation of a B.1.351 SARS-CoV-2 stock on Calu-3 cells eliminated viruses that previously accumulated mutations in the furin cleavage site. Notably, there were alternative variants that accumulated at the same nucleotide positions in virus populations grown on Calu-3 cells at multiple independent facilities. When a Calu-3-derived B.1.351 virus stock was used to infect hamsters, the virus remained pathogenic and the Calu-3-specific variants persisted in the population. These results suggest that Calu-3-derived virus stocks are pathogenic but care should still be taken to evaluate virus stocks for newly arising mutations during propagation. MDPI 2021-12-04 /pmc/articles/PMC8704555/ /pubmed/34960703 http://dx.doi.org/10.3390/v13122434 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Baczenas, John James
Andersen, Hanne
Rashid, Sujatha
Yarmosh, David
Puthuveetil, Nikhita
Parker, Michael
Bradford, Rebecca
Florence, Clint
Stemple, Kimberly J.
Lewis, Mark G.
O’Connor, Shelby L.
Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title_full Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title_fullStr Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title_full_unstemmed Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title_short Propagation of SARS-CoV-2 in Calu-3 Cells to Eliminate Mutations in the Furin Cleavage Site of Spike
title_sort propagation of sars-cov-2 in calu-3 cells to eliminate mutations in the furin cleavage site of spike
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8704555/
https://www.ncbi.nlm.nih.gov/pubmed/34960703
http://dx.doi.org/10.3390/v13122434
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