A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant

Sudhakar Agnihothram, Boyd L. Yount, Eric F. Donaldson, Jeremy Huynh, Vineet Menachery, Lisa E. Gralinski, Rachel L. Graham, Michelle M. Becker, Sakshi Tomar, Trevor D. Scobey, Heather L. Osswald, Alan Whitmore, Robin Gopal, Arun K. Ghosh, Andrew Mesecar, Maria Zambon, Mark Heise, Mark R. Denison, Ralph S. Baric

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Cross-species transmission of zoonotic coronaviruses (CoVs) can result in pandemic disease outbreaks. Middle East respiratory syndrome CoV (MERS-CoV), identified in 2012, has caused 182 cases to date, with ~43% mortality, and no small animal model has been reported. MERS-CoV and Pipistrellus bat coronavirus (BtCoV) strain HKU5 of Betacoronavirus (β-CoV) subgroup 2c share >65% identity at the amino acid level in several regions, including nonstructural protein 5 (nsp5) and the nucleocapsid (N) protein, which are significant drug and vaccine targets. BtCoV HKU5 has been described in silico but has not been shown to replicate in culture, thus hampering drug and vaccine studies against subgroup 2c β-CoVs. We report the synthetic reconstruction and testing of BtCoV HKU5 containing the severe acute respiratory syndrome (SARS)-CoV spike (S) glycoprotein ectodomain (BtCoV HKU5-SE). This virus replicates efficiently in cell culture and in young and aged mice, where the virus targets airway and alveolar epithelial cells. Unlike some subgroup 2b SARS-CoV vaccines that elicit a strong eosinophilia following challenge, we demonstrate that BtCoV HKU5 and MERS-CoV N-expressing Venezuelan equine encephalitis virus replicon particle (VRP) vaccines do not cause extensive eosinophilia following BtCoV HKU5-SE challenge. Passage of BtCoV HKU5-SE in young mice resulted in enhanced virulence, causing 20% weight loss, diffuse alveolar damage, and hyaline membrane formation in aged mice. Passaged virus was characterized by mutations in the nsp13, nsp14, open reading frame 5 (ORF5) and M genes. Finally, we identified an inhibitor active against the nsp5 proteases of subgroup 2c β-CoVs. Synthetic-genome platforms capable of reconstituting emerging zoonotic viral pathogens or their phylogenetic relatives provide new strategies for identifying broad-based therapeutics, evaluating vaccine outcomes, and studying viral pathogenesis.

Original languageEnglish (US)
Article numbere00047-14
JournalmBio
Volume5
Issue number2
DOIs
StatePublished - Mar 25 2014
Externally publishedYes

Fingerprint

Coronavirus
Vaccines
Coronavirus Infections
Severe Acute Respiratory Syndrome
Zoonoses
Eosinophilia
Viruses
Venezuelan Equine Encephalitis Viruses
Alveolar Epithelial Cells
Nucleocapsid Proteins
Replicon
Hyalin
Pandemics
Pharmaceutical Preparations
Virion
Computer Simulation
Open Reading Frames
Disease Outbreaks
Virulence
Weight Loss

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Agnihothram, S., Yount, B. L., Donaldson, E. F., Huynh, J., Menachery, V., Gralinski, L. E., ... Baric, R. S. (2014). A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. mBio, 5(2), [e00047-14]. https://doi.org/10.1128/mBio.00047-14

A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. / Agnihothram, Sudhakar; Yount, Boyd L.; Donaldson, Eric F.; Huynh, Jeremy; Menachery, Vineet; Gralinski, Lisa E.; Graham, Rachel L.; Becker, Michelle M.; Tomar, Sakshi; Scobey, Trevor D.; Osswald, Heather L.; Whitmore, Alan; Gopal, Robin; Ghosh, Arun K.; Mesecar, Andrew; Zambon, Maria; Heise, Mark; Denison, Mark R.; Baric, Ralph S.

In: mBio, Vol. 5, No. 2, e00047-14, 25.03.2014.

Research output: Contribution to journalArticle

Agnihothram, S, Yount, BL, Donaldson, EF, Huynh, J, Menachery, V, Gralinski, LE, Graham, RL, Becker, MM, Tomar, S, Scobey, TD, Osswald, HL, Whitmore, A, Gopal, R, Ghosh, AK, Mesecar, A, Zambon, M, Heise, M, Denison, MR & Baric, RS 2014, 'A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant', mBio, vol. 5, no. 2, e00047-14. https://doi.org/10.1128/mBio.00047-14
Agnihothram S, Yount BL, Donaldson EF, Huynh J, Menachery V, Gralinski LE et al. A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. mBio. 2014 Mar 25;5(2). e00047-14. https://doi.org/10.1128/mBio.00047-14
Agnihothram, Sudhakar ; Yount, Boyd L. ; Donaldson, Eric F. ; Huynh, Jeremy ; Menachery, Vineet ; Gralinski, Lisa E. ; Graham, Rachel L. ; Becker, Michelle M. ; Tomar, Sakshi ; Scobey, Trevor D. ; Osswald, Heather L. ; Whitmore, Alan ; Gopal, Robin ; Ghosh, Arun K. ; Mesecar, Andrew ; Zambon, Maria ; Heise, Mark ; Denison, Mark R. ; Baric, Ralph S. / A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. In: mBio. 2014 ; Vol. 5, No. 2.
@article{ea80ee013bec4043941a5a4352e147f1,
title = "A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant",
abstract = "Cross-species transmission of zoonotic coronaviruses (CoVs) can result in pandemic disease outbreaks. Middle East respiratory syndrome CoV (MERS-CoV), identified in 2012, has caused 182 cases to date, with ~43{\%} mortality, and no small animal model has been reported. MERS-CoV and Pipistrellus bat coronavirus (BtCoV) strain HKU5 of Betacoronavirus (β-CoV) subgroup 2c share >65{\%} identity at the amino acid level in several regions, including nonstructural protein 5 (nsp5) and the nucleocapsid (N) protein, which are significant drug and vaccine targets. BtCoV HKU5 has been described in silico but has not been shown to replicate in culture, thus hampering drug and vaccine studies against subgroup 2c β-CoVs. We report the synthetic reconstruction and testing of BtCoV HKU5 containing the severe acute respiratory syndrome (SARS)-CoV spike (S) glycoprotein ectodomain (BtCoV HKU5-SE). This virus replicates efficiently in cell culture and in young and aged mice, where the virus targets airway and alveolar epithelial cells. Unlike some subgroup 2b SARS-CoV vaccines that elicit a strong eosinophilia following challenge, we demonstrate that BtCoV HKU5 and MERS-CoV N-expressing Venezuelan equine encephalitis virus replicon particle (VRP) vaccines do not cause extensive eosinophilia following BtCoV HKU5-SE challenge. Passage of BtCoV HKU5-SE in young mice resulted in enhanced virulence, causing 20{\%} weight loss, diffuse alveolar damage, and hyaline membrane formation in aged mice. Passaged virus was characterized by mutations in the nsp13, nsp14, open reading frame 5 (ORF5) and M genes. Finally, we identified an inhibitor active against the nsp5 proteases of subgroup 2c β-CoVs. Synthetic-genome platforms capable of reconstituting emerging zoonotic viral pathogens or their phylogenetic relatives provide new strategies for identifying broad-based therapeutics, evaluating vaccine outcomes, and studying viral pathogenesis.",
author = "Sudhakar Agnihothram and Yount, {Boyd L.} and Donaldson, {Eric F.} and Jeremy Huynh and Vineet Menachery and Gralinski, {Lisa E.} and Graham, {Rachel L.} and Becker, {Michelle M.} and Sakshi Tomar and Scobey, {Trevor D.} and Osswald, {Heather L.} and Alan Whitmore and Robin Gopal and Ghosh, {Arun K.} and Andrew Mesecar and Maria Zambon and Mark Heise and Denison, {Mark R.} and Baric, {Ralph S.}",
year = "2014",
month = "3",
day = "25",
doi = "10.1128/mBio.00047-14",
language = "English (US)",
volume = "5",
journal = "mBio",
issn = "2161-2129",
publisher = "American Society for Microbiology",
number = "2",

}

TY - JOUR

T1 - A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant

AU - Agnihothram, Sudhakar

AU - Yount, Boyd L.

AU - Donaldson, Eric F.

AU - Huynh, Jeremy

AU - Menachery, Vineet

AU - Gralinski, Lisa E.

AU - Graham, Rachel L.

AU - Becker, Michelle M.

AU - Tomar, Sakshi

AU - Scobey, Trevor D.

AU - Osswald, Heather L.

AU - Whitmore, Alan

AU - Gopal, Robin

AU - Ghosh, Arun K.

AU - Mesecar, Andrew

AU - Zambon, Maria

AU - Heise, Mark

AU - Denison, Mark R.

AU - Baric, Ralph S.

PY - 2014/3/25

Y1 - 2014/3/25

N2 - Cross-species transmission of zoonotic coronaviruses (CoVs) can result in pandemic disease outbreaks. Middle East respiratory syndrome CoV (MERS-CoV), identified in 2012, has caused 182 cases to date, with ~43% mortality, and no small animal model has been reported. MERS-CoV and Pipistrellus bat coronavirus (BtCoV) strain HKU5 of Betacoronavirus (β-CoV) subgroup 2c share >65% identity at the amino acid level in several regions, including nonstructural protein 5 (nsp5) and the nucleocapsid (N) protein, which are significant drug and vaccine targets. BtCoV HKU5 has been described in silico but has not been shown to replicate in culture, thus hampering drug and vaccine studies against subgroup 2c β-CoVs. We report the synthetic reconstruction and testing of BtCoV HKU5 containing the severe acute respiratory syndrome (SARS)-CoV spike (S) glycoprotein ectodomain (BtCoV HKU5-SE). This virus replicates efficiently in cell culture and in young and aged mice, where the virus targets airway and alveolar epithelial cells. Unlike some subgroup 2b SARS-CoV vaccines that elicit a strong eosinophilia following challenge, we demonstrate that BtCoV HKU5 and MERS-CoV N-expressing Venezuelan equine encephalitis virus replicon particle (VRP) vaccines do not cause extensive eosinophilia following BtCoV HKU5-SE challenge. Passage of BtCoV HKU5-SE in young mice resulted in enhanced virulence, causing 20% weight loss, diffuse alveolar damage, and hyaline membrane formation in aged mice. Passaged virus was characterized by mutations in the nsp13, nsp14, open reading frame 5 (ORF5) and M genes. Finally, we identified an inhibitor active against the nsp5 proteases of subgroup 2c β-CoVs. Synthetic-genome platforms capable of reconstituting emerging zoonotic viral pathogens or their phylogenetic relatives provide new strategies for identifying broad-based therapeutics, evaluating vaccine outcomes, and studying viral pathogenesis.

AB - Cross-species transmission of zoonotic coronaviruses (CoVs) can result in pandemic disease outbreaks. Middle East respiratory syndrome CoV (MERS-CoV), identified in 2012, has caused 182 cases to date, with ~43% mortality, and no small animal model has been reported. MERS-CoV and Pipistrellus bat coronavirus (BtCoV) strain HKU5 of Betacoronavirus (β-CoV) subgroup 2c share >65% identity at the amino acid level in several regions, including nonstructural protein 5 (nsp5) and the nucleocapsid (N) protein, which are significant drug and vaccine targets. BtCoV HKU5 has been described in silico but has not been shown to replicate in culture, thus hampering drug and vaccine studies against subgroup 2c β-CoVs. We report the synthetic reconstruction and testing of BtCoV HKU5 containing the severe acute respiratory syndrome (SARS)-CoV spike (S) glycoprotein ectodomain (BtCoV HKU5-SE). This virus replicates efficiently in cell culture and in young and aged mice, where the virus targets airway and alveolar epithelial cells. Unlike some subgroup 2b SARS-CoV vaccines that elicit a strong eosinophilia following challenge, we demonstrate that BtCoV HKU5 and MERS-CoV N-expressing Venezuelan equine encephalitis virus replicon particle (VRP) vaccines do not cause extensive eosinophilia following BtCoV HKU5-SE challenge. Passage of BtCoV HKU5-SE in young mice resulted in enhanced virulence, causing 20% weight loss, diffuse alveolar damage, and hyaline membrane formation in aged mice. Passaged virus was characterized by mutations in the nsp13, nsp14, open reading frame 5 (ORF5) and M genes. Finally, we identified an inhibitor active against the nsp5 proteases of subgroup 2c β-CoVs. Synthetic-genome platforms capable of reconstituting emerging zoonotic viral pathogens or their phylogenetic relatives provide new strategies for identifying broad-based therapeutics, evaluating vaccine outcomes, and studying viral pathogenesis.

UR - http://www.scopus.com/inward/record.url?scp=84899747216&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84899747216&partnerID=8YFLogxK

U2 - 10.1128/mBio.00047-14

DO - 10.1128/mBio.00047-14

M3 - Article

VL - 5

JO - mBio

JF - mBio

SN - 2161-2129

IS - 2

M1 - e00047-14

ER -