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

被引:22
作者
Baczenas, John James [1 ]
Andersen, Hanne [2 ]
Rashid, Sujatha [3 ,4 ]
Yarmosh, David [3 ,4 ]
Puthuveetil, Nikhita [3 ,4 ]
Parker, Michael [3 ,4 ]
Bradford, Rebecca [3 ,4 ]
Florence, Clint [5 ]
Stemple, Kimberly J. [5 ]
Lewis, Mark G. [2 ]
O'Connor, Shelby L. [1 ,6 ]
机构
[1] UW Madison, Wisconsin Natl Primate Ctr, Madison, WI 53711 USA
[2] BIOQUAL Inc, Rockville, MD 20852 USA
[3] Amer Type Culture Collect ATCC, 10801 Univ Blvd, Manassas, VA 20110 USA
[4] Biodef & Emerging Infect Res Resources Repository, 10801 Univ Blvd, Manassas, VA 20110 USA
[5] NIAID, Off Biodef Res Resources & Translat Res, Div Microbiol & Infect Dis, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA
[6] UW Madison, Dept Pathol & Lab Med, Madison, WI 53705 USA
来源
VIRUSES-BASEL | 2021年 / 13卷 / 12期
基金
美国国家卫生研究院;
关键词
SARS-CoV-2; polymorphisms; Calu-3; stock preparation; INFECTION; ACE2;
D O I
10.3390/v13122434
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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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页数:16
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