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Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation

Viral pathogens with the potential to cause widespread disruption to human health and society continue to emerge or re-emerge around the world. Research on such viruses often involves high biocontainment laboratories (BSL3 or BSL4), but the development of diagnostics, vaccines and therapeutics often...

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Autores principales: Boytz, RuthMabel, Seitz, Scott, Gaudiano, Emily, Patten, J. J., Keiser, Patrick T., Connor, John H., Sharpe, Arlene H., Davey, Robert A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9866162/
https://www.ncbi.nlm.nih.gov/pubmed/36680083
http://dx.doi.org/10.3390/v15010043
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author Boytz, RuthMabel
Seitz, Scott
Gaudiano, Emily
Patten, J. J.
Keiser, Patrick T.
Connor, John H.
Sharpe, Arlene H.
Davey, Robert A.
author_facet Boytz, RuthMabel
Seitz, Scott
Gaudiano, Emily
Patten, J. J.
Keiser, Patrick T.
Connor, John H.
Sharpe, Arlene H.
Davey, Robert A.
author_sort Boytz, RuthMabel
collection PubMed
description Viral pathogens with the potential to cause widespread disruption to human health and society continue to emerge or re-emerge around the world. Research on such viruses often involves high biocontainment laboratories (BSL3 or BSL4), but the development of diagnostics, vaccines and therapeutics often uses assays that are best performed at lower biocontainment. Reliable inactivation is necessary to allow removal of materials to these spaces and to ensure personnel safety. Here, we validate the use of gamma irradiation to inactivate culture supernatants and pellets of cells infected with a representative member of the Filovirus and Coronavirus families. We show that supernatants and cell pellets containing SARS-CoV-2 are readily inactivated with 1.9 MRad, while Ebola virus requires higher doses of 2.6 MRad for supernatants and 3.8 MRad for pellets. While these doses of radiation inactivate viruses, proinflammatory cytokines that are common markers of virus infection are still detected with low losses. The doses required for virus inactivation of supernatants are in line with previously reported values, but the inactivation of cell pellets has not been previously reported and enables new approaches for analysis of protein-based host responses to infection.
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spelling pubmed-98661622023-01-22 Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation Boytz, RuthMabel Seitz, Scott Gaudiano, Emily Patten, J. J. Keiser, Patrick T. Connor, John H. Sharpe, Arlene H. Davey, Robert A. Viruses Article Viral pathogens with the potential to cause widespread disruption to human health and society continue to emerge or re-emerge around the world. Research on such viruses often involves high biocontainment laboratories (BSL3 or BSL4), but the development of diagnostics, vaccines and therapeutics often uses assays that are best performed at lower biocontainment. Reliable inactivation is necessary to allow removal of materials to these spaces and to ensure personnel safety. Here, we validate the use of gamma irradiation to inactivate culture supernatants and pellets of cells infected with a representative member of the Filovirus and Coronavirus families. We show that supernatants and cell pellets containing SARS-CoV-2 are readily inactivated with 1.9 MRad, while Ebola virus requires higher doses of 2.6 MRad for supernatants and 3.8 MRad for pellets. While these doses of radiation inactivate viruses, proinflammatory cytokines that are common markers of virus infection are still detected with low losses. The doses required for virus inactivation of supernatants are in line with previously reported values, but the inactivation of cell pellets has not been previously reported and enables new approaches for analysis of protein-based host responses to infection. MDPI 2022-12-23 /pmc/articles/PMC9866162/ /pubmed/36680083 http://dx.doi.org/10.3390/v15010043 Text en © 2022 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
Boytz, RuthMabel
Seitz, Scott
Gaudiano, Emily
Patten, J. J.
Keiser, Patrick T.
Connor, John H.
Sharpe, Arlene H.
Davey, Robert A.
Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title_full Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title_fullStr Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title_full_unstemmed Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title_short Inactivation of Ebola Virus and SARS-CoV-2 in Cell Culture Supernatants and Cell Pellets by Gamma Irradiation
title_sort inactivation of ebola virus and sars-cov-2 in cell culture supernatants and cell pellets by gamma irradiation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9866162/
https://www.ncbi.nlm.nih.gov/pubmed/36680083
http://dx.doi.org/10.3390/v15010043
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