Biological characterization of cold-adapted SARS-CoV-2 variants
Introduction. The emergence of new epidemiologically significant variants of SARS-CoV-2 has shifted emphasis to development of a live vaccine, which would be able to provide protection against a wide range of antigenic variants of the virus. The aim of the study was to obtain SARS-CoV-2 variants...
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| Format: | Article |
| Language: | Russian |
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Central Research Institute for Epidemiology
2022-09-01
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| Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
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| Online Access: | https://microbiol.crie.ru/jour/article/viewFile/1304/747 |
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| author | Evgeny B. Faizuloev Ekaterina R. Korchevaya Anastasiia V. Gracheva Roman V. Samoilikov Daria I. Smirnova Olga S. Sokolova Grigory S. Glukhov Andrey V. Moiseenko Irina A. Leneva Firaya G. Nagieva Oksana A. Svitich Vitaly V. Zverev |
| author_facet | Evgeny B. Faizuloev Ekaterina R. Korchevaya Anastasiia V. Gracheva Roman V. Samoilikov Daria I. Smirnova Olga S. Sokolova Grigory S. Glukhov Andrey V. Moiseenko Irina A. Leneva Firaya G. Nagieva Oksana A. Svitich Vitaly V. Zverev |
| author_sort | Evgeny B. Faizuloev |
| collection | DOAJ |
| description | Introduction. The emergence of new epidemiologically significant variants of SARS-CoV-2 has shifted emphasis to development of a live vaccine, which would be able to provide protection against a wide range of antigenic variants of the virus.
The aim of the study was to obtain SARS-CoV-2 variants attenuated through cold adaptation and to provide their biological characterization.
Materials and methods. The Dubrovka laboratory strain of SARS-CoV-2 and its variants were cultured on Vero and Calu-3 cells. The virus quantification was performed by virus titration in Vero cells and by real-time reverse transcription-polymerase chain reaction. SARS-CoV-2 virions were analyzed using transmission electron microscopy. Genome sequences of the virus were identified by nanopore sequencing. The attenuation (att) phenotype of SARS-CoV-2 variants was identified using Syrian hamsters as an animal model for COVID-19.
Results. Cold-adapted (ca) SARS-CoV-2 variants Dubrovka-ca-B4 and Dubrovka-ca-D2 were produced by continued passaging of the Dubrovka strain in the Vero cell culture at the temperature being gradually decreased to 23C and by subsequent cloning. Up to 20 nucleotide substitutions and 18 amino acid substitutions were detected in genomes of ca-variants. Ca-variants, as distinct from the parent Dubrovka strain, actively replicated at 23C, while the Dubrovka-ca-D2 variant had a temperature-sensitive (ts) phenotype (did not replicate at 39C). Ca-variants of the virus replicated poorly at 37C in the Calu-3 human lung cell culture, which, along with the ts-phenotype, can be a marker of virus attenuation for humans. In the intranasally infected Syrian hamsters, ca-variants of the virus demonstrated an attenuation phenotype: they did not cause loss of appetite, fatigue, drowsiness, did not slow down weight gain, replicating much more slowly in the lungs and brain compared to the virulent Dubrovka strain.
Conclusion. The obtained attenuated SARS-CoV-2 ca-variants, Dubrovka-ca-B4 and Dubrovka-ca-D2, should be studied further as candidate vaccine strains for a live attenuated vaccine against COVID-19. |
| first_indexed | 2024-03-11T10:50:37Z |
| format | Article |
| id | doaj.art-11077861b78f48f5874a01ab3ab0511b |
| institution | Directory Open Access Journal |
| issn | 0372-9311 2686-7613 |
| language | Russian |
| last_indexed | 2024-03-11T10:50:37Z |
| publishDate | 2022-09-01 |
| publisher | Central Research Institute for Epidemiology |
| record_format | Article |
| series | Журнал микробиологии, эпидемиологии и иммунобиологии |
| spelling | doaj.art-11077861b78f48f5874a01ab3ab0511b2023-11-13T17:19:42ZrusCentral Research Institute for EpidemiologyЖурнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132022-09-0199439740910.36233/0372-9311-280822Biological characterization of cold-adapted SARS-CoV-2 variantsEvgeny B. Faizuloev0https://orcid.org/0000-0001-7385-5083Ekaterina R. Korchevaya1https://orcid.org/0000-0002-6417-3301Anastasiia V. Gracheva2https://orcid.org/0000-0001-8428-4482Roman V. Samoilikov3https://orcid.org/0000-0001-6405-1390Daria I. Smirnova4https://orcid.org/0000-0001-7325-0834Olga S. Sokolova5https://orcid.org/0000-0003-4678-232XGrigory S. Glukhov6https://orcid.org/0000-0002-9540-8249Andrey V. Moiseenko7https://orcid.org/0000-0003-1112-2356Irina A. Leneva8https://orcid.org/0000-0002-7755-2714Firaya G. Nagieva9https://orcid.org/0000-0001-8204-4899Oksana A. Svitich10https://orcid.org/0000-0003-1757-8389Vitaly V. Zverev11https://orcid.org/0000-0001-5808-2246I.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraLomonosov Moscow State University, Faculty of BiologyLomonosov Moscow State University, Faculty of BiologyLomonosov Moscow State University, Faculty of BiologyI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraI.I. Mechnikov Research Institute of Vaccines and SeraIntroduction. The emergence of new epidemiologically significant variants of SARS-CoV-2 has shifted emphasis to development of a live vaccine, which would be able to provide protection against a wide range of antigenic variants of the virus. The aim of the study was to obtain SARS-CoV-2 variants attenuated through cold adaptation and to provide their biological characterization. Materials and methods. The Dubrovka laboratory strain of SARS-CoV-2 and its variants were cultured on Vero and Calu-3 cells. The virus quantification was performed by virus titration in Vero cells and by real-time reverse transcription-polymerase chain reaction. SARS-CoV-2 virions were analyzed using transmission electron microscopy. Genome sequences of the virus were identified by nanopore sequencing. The attenuation (att) phenotype of SARS-CoV-2 variants was identified using Syrian hamsters as an animal model for COVID-19. Results. Cold-adapted (ca) SARS-CoV-2 variants Dubrovka-ca-B4 and Dubrovka-ca-D2 were produced by continued passaging of the Dubrovka strain in the Vero cell culture at the temperature being gradually decreased to 23C and by subsequent cloning. Up to 20 nucleotide substitutions and 18 amino acid substitutions were detected in genomes of ca-variants. Ca-variants, as distinct from the parent Dubrovka strain, actively replicated at 23C, while the Dubrovka-ca-D2 variant had a temperature-sensitive (ts) phenotype (did not replicate at 39C). Ca-variants of the virus replicated poorly at 37C in the Calu-3 human lung cell culture, which, along with the ts-phenotype, can be a marker of virus attenuation for humans. In the intranasally infected Syrian hamsters, ca-variants of the virus demonstrated an attenuation phenotype: they did not cause loss of appetite, fatigue, drowsiness, did not slow down weight gain, replicating much more slowly in the lungs and brain compared to the virulent Dubrovka strain. Conclusion. The obtained attenuated SARS-CoV-2 ca-variants, Dubrovka-ca-B4 and Dubrovka-ca-D2, should be studied further as candidate vaccine strains for a live attenuated vaccine against COVID-19.https://microbiol.crie.ru/jour/article/viewFile/1304/747sars-cov-2cold-adapted virusattenuated virusvaccine strain |
| spellingShingle | Evgeny B. Faizuloev Ekaterina R. Korchevaya Anastasiia V. Gracheva Roman V. Samoilikov Daria I. Smirnova Olga S. Sokolova Grigory S. Glukhov Andrey V. Moiseenko Irina A. Leneva Firaya G. Nagieva Oksana A. Svitich Vitaly V. Zverev Biological characterization of cold-adapted SARS-CoV-2 variants Журнал микробиологии, эпидемиологии и иммунобиологии sars-cov-2 cold-adapted virus attenuated virus vaccine strain |
| title | Biological characterization of cold-adapted SARS-CoV-2 variants |
| title_full | Biological characterization of cold-adapted SARS-CoV-2 variants |
| title_fullStr | Biological characterization of cold-adapted SARS-CoV-2 variants |
| title_full_unstemmed | Biological characterization of cold-adapted SARS-CoV-2 variants |
| title_short | Biological characterization of cold-adapted SARS-CoV-2 variants |
| title_sort | biological characterization of cold adapted sars cov 2 variants |
| topic | sars-cov-2 cold-adapted virus attenuated virus vaccine strain |
| url | https://microbiol.crie.ru/jour/article/viewFile/1304/747 |
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