首页|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

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  • NSTL
  • Elsevier
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'-beta-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.

Tick-borne encephalitis virusReporter virusNeutralization testAntivirals

Driouich, Jean-Selim、Volf, Jiri、Palus, Martin、de Lamballerie, Xavier、Gould, Ernest A.、Ruzek, Daniel、Haviernik, Jan、Eyer, Ludek、Yoshii, Kentaro、Kobayashi, Shintaro、Cerny, Jiri、Nougairede, Antoine

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Aix Marseille Univ, Unite Virus Emergents, IRD 190, Inserm 1207,IHU Mediterranee Infect, Marseille

Vet Res Inst, Hudcova 70, CZ-62100 Brno, Czech Republic

Nagasaki Univ, Natl Res Ctr Control & Prevent Infect Dis, 1-12-4 Sakarnoto, Nagasaki 8528523, Japan

Hokkaido Univ, Fac Vet Med, Lab Publ Hlth, Sapporo, Hokkaido 0600818, Japan

Czech Univ Life Sci Prague, Fac Trop AgriSci, Prague 16500, Czech Republic

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2021

10.1016/j.antiviral.2020.104968

Antiviral research.

Antiviral research.

ISSN：0166-3542
年,卷(期)：2021.185
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