2. Academic Validation
  3. Development and characterization of recombinant tick-borne encephalitis virus expressing mCherry reporter protein: A new tool for high-throughput screening of antiviral compounds, and neutralizing antibody assays

Development and characterization of recombinant tick-borne encephalitis virus expressing mCherry reporter protein: A new tool for high-throughput screening of antiviral compounds, and neutralizing antibody assays

  • Antiviral Res. 2021 Jan;185:104968. doi: 10.1016/j.antiviral.2020.104968.
Jan Haviernik 1,  Ludek Eyer 2,  Kentaro Yoshii 3,  Shintaro Kobayashi 4,  Jiri Cerny 5,  Antoine Nougairède 6,  Jean-Sélim Driouich 6,  Jiri Volf 7,  Martin Palus 2,  Xavier de Lamballerie 6,  Ernest A Gould 6,  Daniel Ruzek 8
Affiliations

Affiliations

  • 1 Veterinary Research Institute, Hudcova 70, CZ-62100, Brno, Czech Republic; Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic.
  • 2 Veterinary Research Institute, Hudcova 70, CZ-62100, Brno, Czech Republic; Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, Branisovska 31, 37005, Ceske Budejovice, Czech Republic.
  • 3 National Research Center for the Control and Prevention of Infectious Diseases, Nagasaki University, 1-12-4 Sakamoto, Nagasaki City 852-8523, Japan; Laboratory of Public Health, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan.
  • 4 Laboratory of Public Health, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, 060-0818, Japan.
  • 5 Faculty of Tropical AgriSciences, Czech University of Life Sciences in Prague, Prague 165 00, Czech Republic.
  • 6 Unité des Virus Émergents (Aix-Marseille Univ-IRD 190-Inserm 1207-IHU Méditerranée Infection), Marseille, France.
  • 7 Veterinary Research Institute, Hudcova 70, CZ-62100, Brno, Czech Republic.
  • 8 Veterinary Research Institute, Hudcova 70, CZ-62100, Brno, Czech Republic; Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, Branisovska 31, 37005, Ceske Budejovice, Czech Republic. Electronic address: [email protected].
PMID: 33157129 DOI: 10.1016/j.antiviral.2020.104968
Abstract

The Flavivirus, tick-borne encephalitis virus (TBEV) is transmitted by Ixodes spp. ticks and may cause severe and potentially lethal neurological tick-borne encephalitis (TBE) in humans. Studying TBEV requires the use of secondary methodologies to detect the virus in infected cells. To overcome this problem, we rationally designed and constructed a recombinant reporter TBEV that stably expressed the mCherry reporter protein. The resulting TBEV reporter virus (named mCherry-TBEV) and wild-type parental TBEV exhibited similar growth kinetics in cultured cells; however, the mCherry-TBEV virus produced smaller plaques. The magnitude of mCherry expression correlated well with progeny virus production but remained stable over <4 passages in Cell Culture. Using well-characterized Antiviral compounds known to inhibit TBEV, 2'-C-methyladenosine and 2'-deoxy-2'-β-hydroxy-4'-azidocytidine (RO-9187), we demonstrated that mCherry-TBEV is suitable for high-throughput screening of Antiviral drugs. Serum samples from a TBEV-vaccinated human and a TBEV-infected dog were used to evaluate the mCherry-based neutralization test. Collectively, recombinant mCherry-TBEV reporter virus described here provides a powerful tool to facilitate the identification of potential Antiviral agents, and to measure levels of neutralizing Antibodies in human and animal sera.

Keywords

Antivirals; Neutralization test; Reporter virus; Tick-borne encephalitis virus.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-10870
    98.39%, HCV Inhibitor
    HCV