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Genome Replication and Regulation of Viral Gene Expression

Establishment of a Virulent Full-Length cDNA Clone for Type I Feline Coronavirus Strain C3663

Yutaka Terada, Yudai Kuroda, Shigeru Morikawa, Yoshiharu Matsuura, Ken Maeda, Wataru Kamitani
Tom Gallagher, Editor
Yutaka Terada
Laboratory of Clinical Research on Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Osaka, JapanLaboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi, Japan
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Yudai Kuroda
Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi, Japan
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Shigeru Morikawa
Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan
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  • ORCID record for Shigeru Morikawa
Yoshiharu Matsuura
Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan
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Ken Maeda
Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi, JapanDepartment of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan
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Wataru Kamitani
Laboratory of Clinical Research on Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan
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Tom Gallagher
Loyola University Chicago
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DOI: 10.1128/JVI.01208-19
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ABSTRACT

Feline infectious peritonitis (FIP) is one of the most important infectious diseases in cats and is caused by feline coronavirus (FCoV). Tissue culture-adapted type I FCoV shows reduced FIP induction in experimental infections, which complicates the understanding of FIP pathogenesis caused by type I FCoV. We previously found that the type I FCoV strain C3663 efficiently induces FIP in specific-pathogen-free cats through the naturally infectious route. In this study, we employed a bacterial artificial chromosome-based reverse genetics system to gain more insights into FIP caused by the C3633 strain. We successfully generated recombinant virus (rC3663) from Fcwf-4 cells transfected with infectious cDNA that showed growth kinetics similar to those shown by the parental virus. Next, we constructed a reporter C3663 virus carrying the nanoluciferase (Nluc) gene to measure viral replication with high sensitivity. The inhibitory effects of different compounds against rC3663-Nluc could be measured within 24 h postinfection. Furthermore, we found that A72 cells derived from canine fibroblasts permitted FCoV replication without apparent cytopathic effects. Thus, our reporter virus is useful for uncovering the infectivity of type I FCoV in different cell lines, including canine-derived cells. Surprisingly, we uncovered aberrant viral RNA transcription of rC3663 in A72 cells. Overall, we succeeded in obtaining infectious cDNA clones derived from type I FCoV that retained its virulence. Our recombinant FCoVs are powerful tools for increasing our understanding of the viral life cycle and pathogenesis of FIP-inducing type I FCoV.

IMPORTANCE Feline coronavirus (FCoV) is one of the most significant coronaviruses, because this virus induces feline infectious peritonitis (FIP), which is a lethal disease in cats. Tissue culture-adapted type I FCoV often loses pathogenicity, which complicates research on type I FCoV-induced feline infectious peritonitis (FIP). Since we previously found that type I FCoV strain C3663 efficiently induces FIP in specific-pathogen-free cats, we established a reverse genetics system for the C3663 strain to obtain recombinant viruses in the present study. By using a reporter C3663 virus, we were able to examine the inhibitory effect of 68 compounds on C3663 replication in Fcwf-4 cells and infectivity in a canine-derived cell line. Interestingly, one canine cell line, A72, permitted FCoV replication but with low efficiency and aberrant viral gene expression.

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Establishment of a Virulent Full-Length cDNA Clone for Type I Feline Coronavirus Strain C3663
Yutaka Terada, Yudai Kuroda, Shigeru Morikawa, Yoshiharu Matsuura, Ken Maeda, Wataru Kamitani
Journal of Virology Oct 2019, 93 (21) e01208-19; DOI: 10.1128/JVI.01208-19

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Establishment of a Virulent Full-Length cDNA Clone for Type I Feline Coronavirus Strain C3663
Yutaka Terada, Yudai Kuroda, Shigeru Morikawa, Yoshiharu Matsuura, Ken Maeda, Wataru Kamitani
Journal of Virology Oct 2019, 93 (21) e01208-19; DOI: 10.1128/JVI.01208-19
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KEYWORDS

coronavirus
viral replication

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