Cargando…

A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection

Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in humans throughout Asia. In the past twenty years, the emergence of the genotype I (GI) JEV as the dominant genotype in Asian countries has raised a significant threat to public health security. However, no clinically appro...

Descripción completa

Detalles Bibliográficos
Autores principales: Li, Chenxi, Chen, Xuan, Hu, Jingbo, Jiang, Daoyuan, Cai, Demin, Li, Yanhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9778660/
https://www.ncbi.nlm.nih.gov/pubmed/36555192
http://dx.doi.org/10.3390/ijms232415548
_version_ 1784856417651392512
author Li, Chenxi
Chen, Xuan
Hu, Jingbo
Jiang, Daoyuan
Cai, Demin
Li, Yanhua
author_facet Li, Chenxi
Chen, Xuan
Hu, Jingbo
Jiang, Daoyuan
Cai, Demin
Li, Yanhua
author_sort Li, Chenxi
collection PubMed
description Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in humans throughout Asia. In the past twenty years, the emergence of the genotype I (GI) JEV as the dominant genotype in Asian countries has raised a significant threat to public health security. However, no clinically approved drug is available for the specific treatment of JEV infection, and the commercial vaccines derived from the genotype III JEV strains merely provided partial protection against the GI JEV. Thus, an easy-to-perform platform in high-throughput is urgently needed for the antiviral drug screening and assessment of neutralizing antibodies specific against the GI JEV. In this study, we established a reverse genetics system for the GI JEV strain (YZ-1) using a homologous recombination strategy. Using this reverse genetic system, a gaussia luciferase (Gluc) expression cassette was inserted into the JEV genome to generate a reporter virus (rGI-Gluc). The reporter virus exhibited similar growth kinetics to the parental virus and remained genetically stable for at least ten passages in vitro. Of note, the bioluminescence signal strength of Gluc in the culture supernatants was well correlated with the viral progenies determined by viral titration. Taking advantage of this reporter virus, we established Gluc readout-based assays for antiviral drug screening and neutralizing antibody detection against the GI JEV. These Gluc readout-based assays exhibited comparable performance to the assays using an actual virus and are less time consuming and are applicable for a high-throughput format. Taken together, we generated a GI JEV reporter virus expressing a Gluc gene that could be a valuable tool for an antiviral drug screening assay and neutralization assay.
format Online
Article
Text
id pubmed-9778660
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-97786602022-12-23 A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection Li, Chenxi Chen, Xuan Hu, Jingbo Jiang, Daoyuan Cai, Demin Li, Yanhua Int J Mol Sci Article Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in humans throughout Asia. In the past twenty years, the emergence of the genotype I (GI) JEV as the dominant genotype in Asian countries has raised a significant threat to public health security. However, no clinically approved drug is available for the specific treatment of JEV infection, and the commercial vaccines derived from the genotype III JEV strains merely provided partial protection against the GI JEV. Thus, an easy-to-perform platform in high-throughput is urgently needed for the antiviral drug screening and assessment of neutralizing antibodies specific against the GI JEV. In this study, we established a reverse genetics system for the GI JEV strain (YZ-1) using a homologous recombination strategy. Using this reverse genetic system, a gaussia luciferase (Gluc) expression cassette was inserted into the JEV genome to generate a reporter virus (rGI-Gluc). The reporter virus exhibited similar growth kinetics to the parental virus and remained genetically stable for at least ten passages in vitro. Of note, the bioluminescence signal strength of Gluc in the culture supernatants was well correlated with the viral progenies determined by viral titration. Taking advantage of this reporter virus, we established Gluc readout-based assays for antiviral drug screening and neutralizing antibody detection against the GI JEV. These Gluc readout-based assays exhibited comparable performance to the assays using an actual virus and are less time consuming and are applicable for a high-throughput format. Taken together, we generated a GI JEV reporter virus expressing a Gluc gene that could be a valuable tool for an antiviral drug screening assay and neutralization assay. MDPI 2022-12-08 /pmc/articles/PMC9778660/ /pubmed/36555192 http://dx.doi.org/10.3390/ijms232415548 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Li, Chenxi
Chen, Xuan
Hu, Jingbo
Jiang, Daoyuan
Cai, Demin
Li, Yanhua
A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title_full A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title_fullStr A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title_full_unstemmed A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title_short A Recombinant Genotype I Japanese Encephalitis Virus Expressing a Gaussia Luciferase Gene for Antiviral Drug Screening Assay and Neutralizing Antibodies Detection
title_sort recombinant genotype i japanese encephalitis virus expressing a gaussia luciferase gene for antiviral drug screening assay and neutralizing antibodies detection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9778660/
https://www.ncbi.nlm.nih.gov/pubmed/36555192
http://dx.doi.org/10.3390/ijms232415548
work_keys_str_mv AT lichenxi arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT chenxuan arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT hujingbo arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT jiangdaoyuan arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT caidemin arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT liyanhua arecombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT lichenxi recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT chenxuan recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT hujingbo recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT jiangdaoyuan recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT caidemin recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection
AT liyanhua recombinantgenotypeijapaneseencephalitisvirusexpressingagaussialuciferasegeneforantiviraldrugscreeningassayandneutralizingantibodiesdetection