Construction and Characterization of Severe Fever with Thrombocytopenia Syndrome Virus with a Fluorescent Reporter for Antiviral Drug Screening

Xiao Wang; Mingyue Xu; Huanhuan Ke; Longda Ma; Liushuai Li; Jiang Li; Fei Deng; Manli Wang; Zhihong Hu; Jia Liu

doi:10.3390/v15051147

Construction and Characterization of Severe Fever with Thrombocytopenia Syndrome Virus with a Fluorescent Reporter for Antiviral Drug Screening

Viruses. 2023 May 10;15(5):1147. doi: 10.3390/v15051147.

Authors

Xiao Wang^{1

2}, Mingyue Xu², Huanhuan Ke², Longda Ma^{2

3}, Liushuai Li², Jiang Li², Fei Deng², Manli Wang^{2

4}, Zhihong Hu^{1

2}, Jia Liu²

Affiliations

¹ Division of Life Science and Medicine, University of Science and Technology of China, Hefei 230027, China.
² State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China.
³ Department of Forensic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
⁴ Hubei Jiangxia Laboratory, Wuhan 430200, China.

Abstract

Severe fever with thrombocytopenia syndrome (SFTS) caused by a novel bunyavirus (SFTSV) is an emerging infectious disease with up to 30% case fatality. Currently, there are no specific antiviral drugs or vaccines for SFTS. Here, we constructed a reporter SFTSV in which the virulent factor nonstructural protein (NSs) was replaced by eGFP for drug screening. First, we developed a reverse genetics system based on the SFTSV HBMC5 strain. Then, the reporter virus SFTSV-delNSs-eGFP was constructed, rescued, and characterized in vitro. SFTSV-delNSs-eGFP showed similar growth kinetics with the wild-type virus in Vero cells. We further detected the antiviral efficacy of favipiravir and chloroquine against wild-type and recombinant SFTSV by the quantification of viral RNA, and compared the results with that of fluorescent assay using high-content screening. The results showed that SFTSV-delNSs-eGFP could be used as a reporter virus for antiviral drug screening in vitro. In addition, we analyzed the pathogenesis of SFTSV-delNSs-eGFP in interferon receptor-deficient (IFNAR^-/-) C57BL/6J mice and found that unlike the fatal infection of the wild-type virus, no obvious pathological change or viral replication were observed in SFTSV-delNSs-eGFP-infected mice. Taken together, the green fluorescence and attenuated pathogenicity make SFTSV-delNSs-eGFP a potent tool for the future high-throughput screening of antiviral drugs.

Keywords: NSs deletion; attenuated virulence; fluorescent virus; high-content antiviral drugs screening; severe fever with thrombocytopenia syndrome virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antiviral Agents / therapeutic use
Bunyaviridae Infections*
Chlorocebus aethiops
Drug Evaluation, Preclinical
Mice
Mice, Inbred C57BL
Phlebovirus*
Severe Fever with Thrombocytopenia Syndrome*
Vero Cells

Substances

Antiviral Agents

Grants and funding

This work was supported by grants from the National Key Research and Development Program of China (2022YFC2303300), the Key Biosafety Science and Technology Program of Hubei Jiangxia Laboratory (JXBS001), the National Natural Science Foundation of China (31900146), and the Hubei Natural Science Foundation for Distinguished Young Scholars (2021CFA050).