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

• Published in: Antiviral research Vol. 185; p. 104968
• Main Authors: Haviernik, Jan, Eyer, Ludek, Yoshii, Kentaro, Kobayashi, Shintaro, Cerny, Jiri, Nougairède, Antoine, Driouich, Jean-Sélim, Volf, Jiri, Palus, Martin, de Lamballerie, Xavier, Gould, Ernest A., Ruzek, Daniel
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2021
• Subjects: Animals; Antibodies, Neutralizing - blood; Antibodies, Viral - blood; Antiviral Agents - isolation & purification; Antivirals; Cell Line; Cricetinae; Encephalitis Viruses, Tick-Borne - genetics; Encephalitis Viruses, Tick-Borne - growth & development; Encephalitis, Tick-Borne - immunology; Encephalitis, Tick-Borne - virology; High-Throughput Screening Assays - methods; Humans; Kidney - cytology; Luminescent Proteins - genetics; Neutralization test; Neutralization Tests; Red Fluorescent Protein; Reporter virus; Tick-borne encephalitis virus; Tick-borne encephalitis virus; Reporter virus; Neutralization test; Antivirals
• DOI: 10.1016/j.antiviral.2020.104968
• ISSN: 0166-3542; 1872-9096

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. •The mCherry-TBEV reporter virus was established.•mCherry-TBEV and wild-type TBEV exhibit similar growth kinetics in cultured cells.•mCherry-TBEV is suitable for high-throughput screening of antiviral drugs.•mCherry-TBEV can be used to measure levels of neutralizing antibodies in human and animal sera.