Chimeric flavivirus between Binjari virus and Murray Valley encephalitis virus
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Flavivirus
West Nile virus
Flavivirus
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Flavivirus
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Togaviridae
Flavivirus
Virus classification
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Mosquito-borne dengue (DEN) and Japanese encephalitis (JE) viruses are the leading causes of arthropod-transmitted viral disease in humans. A licensed tetravalent vaccine that provides effective, long-term immunity against all four serotypes of DEN virus is needed, but is currently unavailable. Improvements to currently available JE vaccines are also needed. Past and recent strategies for the development of new DEN and JE vaccines include inactivated and live attenuated viruses, engineered viruses and chimeric viruses derived from infectious cDNA clones of DEN or JE virus, recombinant poxviruses, recombinant baculoviruses, protein expression in <i>Escherichia coli</i>, and naked DNA vaccines. This report summarizes some of the recent developments in DEN and JE vaccinology, particularly vaccine strategies that involve live attenuated viruses, engineered viruses derived from infectious cDNA clones, and naked DNA vaccines.
Dengue vaccine
Flavivirus
Attenuated vaccine
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Background Japanese encephalitis virus (JEV) is a member of the genus Flavivirus, family Flaviviridae. Flavivirus NS1 glycoproteins are essential proteins, which exhibit a high degree of sequence homology. The NS1 protein is secreted from infected cells as a soluble hexamer and can induce protective immune response in mice against lethal encephalitis. However, its role in neuroinvasion of JEV remains unknown. Our aim is to identify the role of NS1 protein in JEV neuropathogenesis. In this study, we have produced recombinant JEV NS1 protein in Drosophila S2 cells and have characterized its biochemical, biological and antigenic properties.
Flavivirus
Random hexamer
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We have previously reported a system for packaging tick-borne encephalitis (TBE) virus subgenomic replicon RNAs into single-round infectious virus-like particles (VLPs) by using in trans expression of viral C/prM/E structural proteins. In this study, the trans-packaging system was applied to the generation of chimeric VLPs with mosquito-borne Japanese encephalitis (JE) virus. Although trans-expression of TBE virus C and JE virus prM/E proteins resulted in the secretion of VLPs, the expression of JE virus C/prM/E proteins did not lead to the secretion of VLPs, suggesting that homologous interaction between C and non-structural proteins or the genomic RNA is important for efficient assembly of infectious particles. Neutralization testing showed that the antigenic characteristics of the VLPs were similar to those of the native virus. Furthermore, the infectivities of the TBE virus- and JE virus-enveloped VLPs for the ISE6 tick cell line and C6/36 mosquito cell line were investigated. The VLPs were able to enter only those cells that were derived from the natural vectors for the respective viruses. TBE virus replicon RNA packaged in VLPs produced TBE virus non-structural proteins in tick cells, but could neither replicate nor produce viral proteins in mosquito cells. These findings indicate the importance of specific cellular factors for virus entry and replication during flavivirus infection of arthropods. These results demonstrate that chimeric VLPs are useful tools for the study of viral genome packaging and cellular factors involved in vector specificity, with the additional safety aspect that these chimeric VLPs can be used instead of full-length chimeric viruses.
Replicon
Tick-borne encephalitis virus
Flavivirus
Virus-like particle
Semliki Forest virus
Subgenomic mRNA
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Flavivirus
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ABSTRACT A chimeric flavivirus infectious cDNA was constructed by exchanging the premembrane (prM) and envelope (E) genes of the yellow fever virus vaccine strain 17D (YF17D) with the corresponding genes of Modoc virus (MOD). This latter virus belongs to the cluster of the “not-known vector” flaviviruses and is, unlike YF17D, neuroinvasive in SCID mice. Replication of in vitro-transcribed RNA from this chimeric flavivirus was shown by [ 3 H]uridine labeling and RNA analysis. Expression of the MOD prM and E proteins was monitored by radioimmunoprecipitation and revealed that the MOD proteins were correctly and efficiently produced from the chimeric precursor protein. The MOD E protein was shown to be N-linked glycosylated, whereas prM, as predicted from the genome sequence, did not contain N-linked carbohydrates. In Vero cells, the chimeric virus replicated with a similar efficiency as the parental viruses, although it formed smaller plaques than YF17D and MOD. In SCID mice that had been infected intraperitoneally with the chimeric virus, the viral load increased steadily until death. The MOD/YF virus, like MOD from which it had acquired the prM and E structural proteins, but unlike YF, proved neuroinvasive in SCID mice. Animals developed neurological symptoms about 15 days after inoculation and died shortly thereafter. The distribution of MOD/YF RNA in the brain of infected mice was similar to that observed in MOD-infected mice. The observations provide compelling evidence that the determinants of neuroinvasiveness of flaviviruses are entirely located in the envelope proteins prM and E.
Flavivirus
Vero cell
Togaviridae
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Alphavirus is a genus of arthropod-borne viruses (arboviruses) in the family Togaviridae (Calisher et al., 1980; Calisher & Karabatsos, 1988). These viruses have unsegmented, single-stranded RNA genomes of 11–12 kb which are of positive or messenger sense. Alphavirus genomic RNA encodes four non-structural proteins, designated nsP1–4, as well as three structural proteins: the capsid, and two envelope glycoproteins, El and E2 (Fig. 33.1). A sub-genomic 26S RNA species, which encodes only the structural proteins, is also produced in infected cells.
Togaviridae
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