A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

npj Vaccines, Journal Year: 2021, Volume and Issue: 6(1)

Published: June 17, 2021

Abstract An array of SARS-CoV-2 virus variants have been isolated, propagated and used in vitro assays, vivo animal studies human clinical trials. Observations working stocks suggest that sequential propagation Vero cells leads to critical changes the region furin cleavage site, which significantly reduce value stock for research studies. Serially propagating E6 rapid increases genetic while other cell lines (e.g. Vero/hSLAM) appears mitigate this risk thereby improving overall stability stocks. From these observations, investigators are urged monitor carefully when cells.

Language: Английский

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First evidence that an emerging mammalian alphacoronavirus is able to infect an avian species

Transboundary and Emerging Diseases, Journal Year: 2022, Volume and Issue: 69(5)

Published: March 27, 2022

A novel swine enteric alphacoronavirus, acute diarrhoea syndrome coronavirus (SADS-CoV), related to Rhinolophus bat CoV HKU2 in the subgenus Rhinacovirus emerged southern China 2017, causing newborn piglets, and critical questions remain about pathogenicity, cross-species transmission potential animal reservoirs. Our laboratory's previous research has shown that SADS-CoV can replicate various cell types from different species, including chickens. Here, we systematically explore susceptibility of chickens a cell-adapted strain both vitro vivo. First, evidence replication primary chicken cells, cytopathic effects, immunofluorescence staining, growth curves structural protein expression, was proven. Furthermore, observed replicated embryos without gross lesions experimental infection chicks resulted mild respiratory symptoms. More importantly, shedding viral distribution lungs, spleens, small intestines large infected were confirmed by quantitative reverse transcription polymerase chain reaction immunohistochemistry. The genomic sequence original pig source sample 2017 determined have nine nucleotide differences compared used; among these three nonsynonymous mutations spike gene. These results collectively demonstrate are susceptible infection, suggesting they reservoir. To our knowledge, this study provides first which mammalian alphacoronavirus is able infect an avian species.

Language: Английский

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Diversity of Coronaviruses with Particular Attention to the Interspecies Transmission of SARS-CoV-2

Animals, Journal Year: 2022, Volume and Issue: 12(3), P. 378 - 378

Published: Feb. 4, 2022

In December 2019, the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was reported in China with serious impacts on global health and economy that is still ongoing. Although interspecies transmission coronaviruses common well documented, each has a narrowly restricted host range. Coronaviruses utilize different receptors to mediate membrane fusion replication cell cytoplasm. The interplay between receptor-binding domain (RBD) their coevolution are determinants for susceptibility. recently emerged SARS-CoV-2 caused disease 2019 (COVID-19) pandemic also been domestic wild animals, raising question about responsibility animals virus evolution. Additionally, COVID-19 might substantially have an impact animal production long time. present review, we discussed diversity thus receptors. Moreover, susceptibility several special reference current evidence were highlighted. Finally, shed light urgent demand implementation One Health concept as collaborative approach mitigate threat both humans animals.

Language: Английский

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Cell culture systems for isolation of SARS-CoV-2 clinical isolates and generation of recombinant virus

iScience, Journal Year: 2023, Volume and Issue: 26(5), P. 106634 - 106634

Published: April 10, 2023

A simple and robust cell culture system is essential for generating authentic SARS-CoV-2 stocks evaluation of viral pathogenicity, screening antiviral compounds, preparation inactivated vaccines. Evidence suggests that Vero E6, a line commonly used in the field to grow SARS-CoV-2, does not support efficient propagation new variants triggers rapid adaptation virus. We generated panel 17 human lines overexpressing entry factors tested their ability infection. Two lines, Caco-2/AT HuH-6/AT, demonstrated exceptional susceptibility, yielding highly concentrated virus stocks. Notably, these were more sensitive than E6 cells recovering from clinical specimens. Further, provided platform producing genetically reliable recombinant through reverse genetics system. These cellular models are valuable tool study its continuously emerging variants.

Language: Английский

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Systematic Exploration of SARS-CoV-2 Adaptation to Vero E6, Vero E6/TMPRSS2, and Calu-3 Cells

Genome Biology and Evolution, Journal Year: 2023, Volume and Issue: 15(4)

Published: Feb. 28, 2023

Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread globally, and scientists around the world are currently studying virus intensively in order fight against on-going pandemic of virus. To do so, SARS-CoV-2 is typically grown lab generate viral stocks for various kinds experimental investigations. However, accumulating evidence suggests that such viruses often undergo cell culture adaptation. Here, we systematically explored adaptation two variants, namely B.1.36.16 variant AY.30 variant, a sub lineage B.1.617.2 (Delta) propagated three different lines, including Vero E6, E6/TMPRSS2, Calu-3 cells. Our analyses detected numerous potential changes scattering across entire genome, many which could be found naturally circulating isolates. Notable ones included mutations spike glycoprotein's multibasic cleavage site, Omicron-defining H655Y mutation on glycoprotein, as well nucleocapsid protein's linker region, all were E6-specific. also identified deletion non-structural protein 1 membrane glycoprotein Calu-3-specific changes. S848C 3, located papain-like protease domain, was change, lines. results highlight high adaptability, emphasize need deep-sequence cultured samples when used intricate sensitive biological experiments, illustrate power evolutionary study shedding lights landscape.

Language: Английский

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