Construction of an infectious molecular clone of Japanese encephalitis virus genotype V and its derivative subgenomic replicon capable of expressing a foreign gene

Ishikawa, Takahiro; Abe, Makoto; Masuda, Michiaki

doi:10.1016/j.virusres.2014.10.010

Cited by 11 publications

(19 citation statements)

References 59 publications

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“…To clarify the regions responsible for increased pathogenicity in the Muar genome, various GI-GV intertypic and point mutant viruses may be required. Recently, Ishikawa et al (2015) identified the infectious molecular clone and reporter replicon of the Muar strain, which is also beneficial for generally understanding the characteristics of GV JEV. In addition, de Wispelaere et al (2015) constructed a molecular clone of the GV XZ0934 JEV strain and demonstrated that this recombinant GV virus exhibited a greater pathogenicity in BALB/c mice than that of GIII JEV.…”

Section: Discussionmentioning

confidence: 99%

In vitro growth, pathogenicity and serological characteristics of the Japanese encephalitis virus genotype V Muar strain

Tajima

Yagasaki

Kotaki

et al. 2015

Journal of General Virology

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The characteristics of genotype V Japanese encephalitis virus (GV JEV) remain poorly understood as only two strains have been isolated to date. In this study, we examined the effects of the GV JEV Muar strain on in vitro growth and pathogenicity in mice; we also evaluated the efficacy of inactivated JEV vaccines against the Muar strain. Although growth of the Muar strain in mouse neuroblastoma N18 cells was clearly worse than that of the GIII Beijing-1 and GI Mie/41/2002 strains, neuroinvasiveness of the Muar strain was similar to that of the Beijing-1 strain and significantly higher than that of the Mie/41/2002 strain. The results of a plaque reduction neutralization test suggested that the neutralization ability of the JEV vaccines against the Muar strain was reduced compared with the GI and GIII strains. However, the protection potency of the JEV vaccine against the Muar strain was similar to that for the Beijing-1 strain in mice. Our data indicate that GV JEV has unique growth, virulence and antigenicity features.

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Section: Discussionmentioning

confidence: 99%

In vitro growth, pathogenicity and serological characteristics of the Japanese encephalitis virus genotype V Muar strain

Tajima

Yagasaki

Kotaki

et al. 2015

Journal of General Virology

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“…Unfortunately, although most reverse genetics systems proved to be efficient to recover arboviruses from vertebrate cell lines (for reviews see 4 , 5 ), very few studies have reported such systems for arthropod cells and especially for cells from Aedes mosquitoes, one of the most important arbovirus vectors globally 6 . Indeed, reverse genetics systems designed for positive-sense single-stranded RNA viruses in Aedes mosquito cells are typically based to date on the lipofection or electroporation of synthetic capped RNA transcripts generated by in vitro transcription from SP6- 7 – 9 or T7 promoter-driven 10 – 16 full-length viral cDNA constructs. A second system only used marginally and based on the direct transfection of a T7 promoter-driven infectious clone in an Aedes mosquito cell line stably expressing the T7 RNA polymerase was established to produce a minireplicon of the Bunyamwera negative-strand RNA virus 17 .…”

Section: Introductionmentioning

confidence: 99%

New reverse genetics and transfection methods to rescue arboviruses in mosquito cells

Atieh

Nougairède

Klitting

et al. 2017

Sci Rep

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Reverse genetics is a critical tool to decrypt the biological properties of arboviruses. However, whilst reverse genetics methods have been usually applied to vertebrate cells, their use in insect cells remains uncommon due to the conjunction of laborious molecular biology techniques and of specific difficulties surrounding the transfection of such cells. To leverage reverse genetics studies in both vertebrate and mosquito cells, we designed an improved DNA transfection protocol for insect cells and then demonstrated that the simple and flexible ISA (Infectious Subgenomic Amplicons) reverse-genetics method can be efficiently applied to both mammalian and mosquito cells to generate in days recombinant infectious positive-stranded RNA viruses belonging to genera Flavivirus (Japanese encephalitis, Yellow fever, West Nile and Zika viruses) and Alphavirus (Chikungunya virus). This method represents an effective option to potentially overcome technological issues related to the study of arboviruses.

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“…A recent report described the construction of molecular virology tools that will help characterize the prototype strain Muar (15). In the present study, we used a cDNA-based technology to produce a g5 virus derived from the recently isolated strain XZ0934 (9).…”

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confidence: 99%

A Japanese Encephalitis Virus Genotype 5 Molecular Clone Is Highly Neuropathogenic in a Mouse Model: Impact of the Structural Protein Region on Virulence

Wispelaere

Frenkiel

Desprès

2015

J Virol

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Japanese encephalitis virus (JEV) strains can be separated into 5 genotypes (g1 to g5) based on sequence similarity. JEV g5 strains have been rarely isolated and are poorly characterized. We report here the full characterization of a g5 virus generated using a cDNA-based technology and its comparison with a widely studied g3 strain. We did not observe any major differences between those viruses when their infectious cycles were studied in various cell lines in vitro. Interestingly, the JEV g5 strain was highly pathogenic when inoculated to BALB/c mice, which are known to be largely resistant to JEV g3 infection. The study of chimeric viruses between JEV g3 and g5 showed that there was a poor viral clearance of viruses that express JEV g5 structural proteins in BALB/c mice blood, which correlated with viral invasion of the central nervous system and encephalitis. In addition, using an in vitro model of the blood-brain barrier, we were able to show that JEV g5 does not have an enhanced capacity for entering the central nervous system, compared to JEV g3. Overall, in addition to providing a first characterization of the understudied JEV g5, our work highlights the importance of sustaining an early viremia in the development of JEV encephalitis. IMPORTANCEGenotype 5 viruses are genetically and serologically distinct from other JEV genotypes and can been associated with human encephalitis, which warrants the need for their characterization. In this study, we characterized the in vitro and in vivo properties of a JEV g5 strain and showed that it was more neuropathogenic in a mouse model than a well-characterized JEV g3 strain. The enhanced virulence of JEV g5 was associated with poor viral clearance but not with enhanced crossing of the blood-brain barrier, thus providing new insights into JEV pathogenesis. JEV is a significant human pathogen and the causative agent for Japanese encephalitis, one of the most important viral encephalitides of medical interest in Asia, with an incidence of approximately 67,900 cases per year, among which are about 20,000 fatal cases (1). About 20 to 30% of the symptomatic human cases are fatal, while 30 to 50% of survivors can develop long-term neurological sequelae (2). JEV is maintained in an enzootic cycle between Culex tritaeniorhynchus mosquitoes and amplifying vertebrate hosts, such as water birds and domestic swine (3). Humans and several other animals can also be infected, but since they do not develop a sufficient level of viremia to infect mosquitoes, they are thought to be dead-end hosts (4).Phylogenetic studies based on the viral envelope protein sequences allow the division of JEV strains into five genotypes (g1 to g5). From the isolation of the prototype strain of JEV in 1935 until recently, most of the circulating strains of JEV belonged to g3 and were at the origin of major epidemics in Southeast Asian countries (5). Recently, a shift in prevalence from JEV g3 to g1 has been observed in several Asian countries (6-8), while some strains of JEV g5 have been occasio...

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Construction of an infectious molecular clone of Japanese encephalitis virus genotype V and its derivative subgenomic replicon capable of expressing a foreign gene

Cited by 11 publications

References 59 publications

In vitro growth, pathogenicity and serological characteristics of the Japanese encephalitis virus genotype V Muar strain

In vitro growth, pathogenicity and serological characteristics of the Japanese encephalitis virus genotype V Muar strain

New reverse genetics and transfection methods to rescue arboviruses in mosquito cells

A Japanese Encephalitis Virus Genotype 5 Molecular Clone Is Highly Neuropathogenic in a Mouse Model: Impact of the Structural Protein Region on Virulence

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