Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis

ABSTRACT Feline infectious peritonitis (FIP), one of the most important lethal infections of cats, is caused by feline infectious peritonitis virus (FIPV), the high-virulence biotype of feline coronaviruses (FCoVs). FIPVs are suggested to emerge from feline enteric coronaviruses (FECVs) by acquiring...

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Autores principales:	Rosina Ehmann, Claudia Kristen-Burmann, Barbara Bank-Wolf, Matthias König, Christiane Herden, Torsten Hain, Heinz-Jürgen Thiel, John Ziebuhr, Gergely Tekes
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	feline coronavirus field isolates feline infectious peritonitis reverse genetics Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/230b734ecb4f465f913b84e1a7f471a9
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spelling	oai:doaj.org-article:230b734ecb4f465f913b84e1a7f471a92021-11-15T16:00:14ZReverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis10.1128/mBio.01422-182150-7511https://doaj.org/article/230b734ecb4f465f913b84e1a7f471a92018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01422-18https://doaj.org/toc/2150-7511ABSTRACT Feline infectious peritonitis (FIP), one of the most important lethal infections of cats, is caused by feline infectious peritonitis virus (FIPV), the high-virulence biotype of feline coronaviruses (FCoVs). FIPVs are suggested to emerge from feline enteric coronaviruses (FECVs) by acquiring mutations in specific genes in the course of persistent infections. Although numerous studies identified mutations predicted to be responsible for the FECV-FIPV biotype switch, the presumed roles of specific genetic changes in FIP pathogenesis have not been confirmed experimentally. Reverse genetics systems established previously for serotype I and the less common serotype II FCoVs were based on cell culture-adapted FIPV strains which, however, were shown to be unsuitable for FIP pathogenesis studies in vivo. To date, systems to produce and manipulate recombinant serotype I field viruses have not been developed, mainly because these viruses cannot be grown in vitro. Here, we report the first reverse genetics system based on a serotype I FECV field isolate that is suitable to produce high-titer stocks of recombinant FECVs. We demonstrate that these recombinant viruses cause productive persistent infections in cats that are similar to what is observed in natural infections. The system provides an excellent tool for studying FCoVs that do not grow in standard cell culture systems and will greatly facilitate studies into the molecular pathogenesis of FIP. Importantly, the system could also be adapted for studies of other RNA viruses with large genomes whose production and characterization in vivo are currently hampered by the lack of in vitro propagation systems. IMPORTANCE The availability of recombinant serotype I FCoV field isolates that are amenable to genetic manipulation is key to studying the molecular pathogenesis of FIP, especially since previous studies using cell culture-adapted FIPVs had proven unsuccessful. To our knowledge, we report the first serotype I FECV field isolate-based reverse genetics system that allows the production of high-titer recombinant virus stocks that can be used for subsequent in vivo studies in cats. The system represents a milestone in FCoV research. It provides an essential tool for studying the molecular pathogenesis of FIP and, more specifically, the functions of specific gene products in causing a fundamentally different progression of disease following acquisition of specific mutations. The system developed in this study will also be useful for studying other coronaviruses or more distantly related RNA viruses with large genomes for which suitable in vitro culture systems are not available.Rosina EhmannClaudia Kristen-BurmannBarbara Bank-WolfMatthias KönigChristiane HerdenTorsten HainHeinz-Jürgen ThielJohn ZiebuhrGergely TekesAmerican Society for Microbiologyarticlefeline coronavirus field isolatesfeline infectious peritonitisreverse geneticsMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	feline coronavirus field isolates feline infectious peritonitis reverse genetics Microbiology QR1-502
spellingShingle	feline coronavirus field isolates feline infectious peritonitis reverse genetics Microbiology QR1-502 Rosina Ehmann Claudia Kristen-Burmann Barbara Bank-Wolf Matthias König Christiane Herden Torsten Hain Heinz-Jürgen Thiel John Ziebuhr Gergely Tekes Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
description	ABSTRACT Feline infectious peritonitis (FIP), one of the most important lethal infections of cats, is caused by feline infectious peritonitis virus (FIPV), the high-virulence biotype of feline coronaviruses (FCoVs). FIPVs are suggested to emerge from feline enteric coronaviruses (FECVs) by acquiring mutations in specific genes in the course of persistent infections. Although numerous studies identified mutations predicted to be responsible for the FECV-FIPV biotype switch, the presumed roles of specific genetic changes in FIP pathogenesis have not been confirmed experimentally. Reverse genetics systems established previously for serotype I and the less common serotype II FCoVs were based on cell culture-adapted FIPV strains which, however, were shown to be unsuitable for FIP pathogenesis studies in vivo. To date, systems to produce and manipulate recombinant serotype I field viruses have not been developed, mainly because these viruses cannot be grown in vitro. Here, we report the first reverse genetics system based on a serotype I FECV field isolate that is suitable to produce high-titer stocks of recombinant FECVs. We demonstrate that these recombinant viruses cause productive persistent infections in cats that are similar to what is observed in natural infections. The system provides an excellent tool for studying FCoVs that do not grow in standard cell culture systems and will greatly facilitate studies into the molecular pathogenesis of FIP. Importantly, the system could also be adapted for studies of other RNA viruses with large genomes whose production and characterization in vivo are currently hampered by the lack of in vitro propagation systems. IMPORTANCE The availability of recombinant serotype I FCoV field isolates that are amenable to genetic manipulation is key to studying the molecular pathogenesis of FIP, especially since previous studies using cell culture-adapted FIPVs had proven unsuccessful. To our knowledge, we report the first serotype I FECV field isolate-based reverse genetics system that allows the production of high-titer recombinant virus stocks that can be used for subsequent in vivo studies in cats. The system represents a milestone in FCoV research. It provides an essential tool for studying the molecular pathogenesis of FIP and, more specifically, the functions of specific gene products in causing a fundamentally different progression of disease following acquisition of specific mutations. The system developed in this study will also be useful for studying other coronaviruses or more distantly related RNA viruses with large genomes for which suitable in vitro culture systems are not available.
format	article
author	Rosina Ehmann Claudia Kristen-Burmann Barbara Bank-Wolf Matthias König Christiane Herden Torsten Hain Heinz-Jürgen Thiel John Ziebuhr Gergely Tekes
author_facet	Rosina Ehmann Claudia Kristen-Burmann Barbara Bank-Wolf Matthias König Christiane Herden Torsten Hain Heinz-Jürgen Thiel John Ziebuhr Gergely Tekes
author_sort	Rosina Ehmann
title	Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
title_short	Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
title_full	Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
title_fullStr	Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
title_full_unstemmed	Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis
title_sort	reverse genetics for type i feline coronavirus field isolate to study the molecular pathogenesis of feline infectious peritonitis
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/230b734ecb4f465f913b84e1a7f471a9
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Reverse Genetics for Type I Feline Coronavirus Field Isolate To Study the Molecular Pathogenesis of Feline Infectious Peritonitis

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