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
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
33157129
DOI
10.1016/j.antiviral.2020.104968
PII: S0166-3542(20)30382-X
Knihovny.cz E-zdroje
- Klíčová slova
- Antivirals, Neutralization test, Reporter virus, Tick-borne encephalitis virus,
- MeSH
- antivirové látky izolace a purifikace MeSH
- buněčné linie MeSH
- červený fluorescenční protein MeSH
- klíšťová encefalitida imunologie virologie MeSH
- křečci praví MeSH
- ledviny cytologie MeSH
- lidé MeSH
- luminescentní proteiny genetika MeSH
- neutralizační testy * MeSH
- neutralizující protilátky krev MeSH
- protilátky virové krev MeSH
- rychlé screeningové testy metody MeSH
- viry klíšťové encefalitidy genetika růst a vývoj MeSH
- zvířata MeSH
- Check Tag
- křečci praví MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antivirové látky MeSH
- luminescentní proteiny MeSH
- neutralizující protilátky MeSH
- protilátky virové MeSH
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.
Unité des Virus Émergents Marseille France
Veterinary Research Institute Hudcova 70 CZ 62100 Brno Czech Republic
Citace poskytuje Crossref.org
