Journal
ANTIVIRAL RESEARCH
Volume 185, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.antiviral.2020.104968
Keywords
Tick-borne encephalitis virus; Reporter virus; Neutralization test; Antivirals
Categories
Funding
- Czech Science Foundation [20-14325 S]
- Czech Academy of Sciences [JSPS-18-08]
- Japanese Society for Promotion of Science
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Researchers developed a recombinant mCherry-expressing reporter virus for TBEV, which enables high-throughput screening of antiviral drugs and evaluation of neutralizing antibodies in serum samples. This reporter virus provides a powerful tool for identifying potential antiviral agents and monitoring immune responses to TBEV in human and animal sera.
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.
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