Development and Characterization of a cDNA-Launch Recombinant Simian Hemorrhagic Fever Virus Expressing Enhanced Green Fluorescent Protein: ORF 2b’ Is Not Required for In Vitro Virus Replication

Yingyun Cai; Shuiqing Yu; Ying Fang; Laura Bollinger; Yanhua Li; Michael Lauck; Elena N. Postnikova; Steven Mazur; Reed F. Johnson; Courtney L. Finch; Sheli R. Radoshitzky; Gustavo Palacios; Thomas C. Friedrich; Tony L. Goldberg; David H. O’Connor; Peter B. Jahrling; Jens H. Kuhn

doi:10.3390/v13040632

Viruses (Apr 2021)

Development and Characterization of a cDNA-Launch Recombinant Simian Hemorrhagic Fever Virus Expressing Enhanced Green Fluorescent Protein: ORF 2b’ Is Not Required for In Vitro Virus Replication

Yingyun Cai,
Shuiqing Yu,
Ying Fang,
Laura Bollinger,
Yanhua Li,
Michael Lauck,
Elena N. Postnikova,
Steven Mazur,
Reed F. Johnson,
Courtney L. Finch,
Sheli R. Radoshitzky,
Gustavo Palacios,
Thomas C. Friedrich,
Tony L. Goldberg,
David H. O’Connor,
Peter B. Jahrling,
Jens H. Kuhn

Affiliations

Yingyun Cai: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Shuiqing Yu: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Ying Fang: College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USA
Laura Bollinger: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Yanhua Li: College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USA
Michael Lauck: Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin–Madison, Madison, WI 53705, USA
Elena N. Postnikova: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Steven Mazur: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Reed F. Johnson: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Courtney L. Finch: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Sheli R. Radoshitzky: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702, USA
Gustavo Palacios: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702, USA
Thomas C. Friedrich: Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin–Madison, Madison, WI 53705, USA
Tony L. Goldberg: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin–Madison, Madison, WI 53706, USA
David H. O’Connor: Department of Pathology and Laboratory Medicine, School of Medicine and Public Health, University of Wisconsin–Madison, Madison, WI 53705, USA
Peter B. Jahrling: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA
Jens H. Kuhn: Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA

DOI: https://doi.org/10.3390/v13040632
Journal volume & issue: Vol. 13, no. 4
p. 632

Abstract

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Simian hemorrhagic fever virus (SHFV) causes acute, lethal disease in macaques. We developed a single-plasmid cDNA-launch infectious clone of SHFV (rSHFV) and modified the clone to rescue an enhanced green fluorescent protein-expressing rSHFV-eGFP that can be used for rapid and quantitative detection of infection. SHFV has a narrow cell tropism in vitro, with only the grivet MA-104 cell line and a few other grivet cell lines being susceptible to virion entry and permissive to infection. Using rSHFV-eGFP, we demonstrate that one cricetid rodent cell line and three ape cell lines also fully support SHFV replication, whereas 55 human cell lines, 11 bat cell lines, and three rodent cells do not. Interestingly, some human and other mammalian cell lines apparently resistant to SHFV infection are permissive after transfection with the rSHFV-eGFP cDNA-launch plasmid. To further demonstrate the investigative potential of the infectious clone system, we introduced stop codons into eight viral open reading frames (ORFs). This approach suggested that at least one ORF, ORF 2b’, is dispensable for SHFV in vitro replication. Our proof-of-principle experiments indicated that rSHFV-eGFP is a useful tool for illuminating the understudied molecular biology of SHFV.

Published in Viruses

ISSN: 1999-4915 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.mdpi.com/journal/viruses

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