Insights into the Pathogenesis and Development of Recombinant Japanese Encephalitis Virus Genotype 3 as a Vaccine

Japanese encephalitis virus (JEV), a flavivirus transmitted by mosquitoes, has caused epidemics and severe neurological diseases in Asian countries. In this study, we developed a cDNA infectious clone, pBAC JYJEV3, of the JEV genotype 3 strain (EF571853.1) using a bacterial artificial chromosome (BA...

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Bibliographic Details
Published in:Vaccines (Basel) 2024-05, Vol.12 (6), p.597
Main Authors: Park, Jae-Yeon, Lee, Hye-Mi, Jun, Sung-Hoon, Kamitani, Wataru, Kim, Onnuri, Shin, Hyun-Jin
Format: Article
Language:English
Subjects:
Antibodies
Aquatic insects
Artificial chromosomes
Bacteria
Bacterial artificial chromosomes
cDNA
Cloning
Comparative analysis
Confocal microscopy
Cytomegalovirus
Design and construction
Disease transmission
E coli
Electron microscopy
Encephalitis
Env protein
Genomes
Genotype & phenotype
Genotypes
Growth kinetics
Hepatitis
Hepatitis delta virus
Immunization
Immunoglobulin G
infectious clone
Infectivity
Japanese encephalitis virus 3
Materials
Microscopy
Morphology
Mosquitoes
Nakayama
Neurological diseases
Pathogenesis
Plasmids
RNA polymerase
Toxicity
Vaccine development
Vaccines
Vector-borne diseases
Vectors (Biology)
Vero cells
Viral envelope proteins
Viruses
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Description
Summary:Japanese encephalitis virus (JEV), a flavivirus transmitted by mosquitoes, has caused epidemics and severe neurological diseases in Asian countries. In this study, we developed a cDNA infectious clone, pBAC JYJEV3, of the JEV genotype 3 strain (EF571853.1) using a bacterial artificial chromosome (BAC) vector. The constructed infectious clone was transfected into Vero cells, where it exhibited infectivity and induced cytopathic effects akin to those of the parent virus. Confocal microscopy confirmed the expression of the JEV envelope protein. Comparative analysis of growth kinetics revealed similar replication dynamics between the parental and recombinant viruses, with peak titers observed 72 h post-infection (hpi). Furthermore, plaque assays demonstrated comparable plaque sizes and morphologies between the viruses. Cryo-electron microscopy confirmed the production of recombinant virus particles with a morphology identical to that of the parent virus. Immunization studies in mice using inactivated parental and recombinant viruses revealed robust IgG responses, with neutralizing antibody production increasing over time. These results showcase the successful generation and characterization of a recombinant JEV3 virus and provide a platform for further investigations into JEV pathogenesis and vaccine development.
ISSN:2076-393X
2076-393X
DOI:10.3390/vaccines12060597