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

0537485 - BC 2022 RIV NL eng J - Journal Article
Haviernik, J. - Eyer, Luděk - Yoshii, K. - Kobayashi, S. - Černý, J. - Nougairède, A. - Driouich, J.-S. - Volf, J. - Palus, Martin - de Lamballerie, X. - Gould, E. A. - Růžek, Daniel
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 Research. Roč. 185, Jan 2021 (2021), č. článku 104968. ISSN 0166-3542. E-ISSN 1872-9096
R&D Projects: GA ČR(CZ) GA20-14325S
Grant - others:AV ČR(CZ) JSPS-18-08
Program: Bilaterální spolupráce
Institutional support: RVO:60077344
Keywords : Antivirals * Neutralization test * Reporter virus * Tick-borne encephalitis virus
OECD category: Microbiology
Impact factor: 10.103, year: 2021
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/pii/S016635422030382X?via%3Dihub

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.
Permanent Link: http://hdl.handle.net/11104/0315299