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Design Of Recombinant Viral Vaccines Based On Mumps Virus

BACKGROUND: Biological platforms such as recombinant viral vectors are particularly interesting for the development ofvaccines against emerging viral diseases or diseases for which vaccines are not yet available. So far, only few studies haveshown the potential of mumps virus (Mumps orthorubulavirus, family Paramyxoviridae) to be used as a recombinant vector.The generation of a recombinant virus expressing HIV-1 Gag protein (Xu R et al., J Virol 83 (2009) 9813) indicated that viralvaccine vectors based on mumps can be produced. OBJECTIVES: The development of recombinant vaccines for hepatitis C virus (HCV) and human respiratory syncytial virus(RSV) based on mumps recombinant virus (vMRV2). Virus vMRV2 was generated using a reverse genetics technology(rescue) and has a genome sequence based on the consensus of vaccine strain L-Zagreb. METHODS: The rescue method we use comprises a co- transfection of BSRT7 cells with three expression plasmids thatencode nucleocapsid protein and components of viral RNA-dependant RNA polymerase (phosphoprotein and large protein), and a plasmid with full-length virus genome. Plasmid pMRV2 contains solely a mumps backbone sequence. It was furthermodified to encode additional genes in order to produce recombinant live attenuated vaccines for HCV and RSV. Sequencesencoding ectodomains of RSV fusion (F) protein and HCV E1 and E2 proteins were cloned into the pMRV2 with or withoutthe addition of transmembrane and cytoplasmic domain (TMD/CD) of mumps F protein. We expect that RSV and HCV geneswill be expressed in infected cells ; proteins with the addition of TMD/CD of mumps F protein will be incorporated in viralparticles as well. RESULTS: Using our reverse genetics system, we have produced recombinant viruses vMRV2, vF RSV- MRV2 (vectorvaccine for RSV) and vE1E2TMD HCV-MRV2 (vector vaccine 1 for HCV) ; rescue for vCE1E2 HCV- MRV2 (vector vaccine2 for HCV) is currently in progress. Viral genetic stability was analysed by next generation sequencing after passaging of vFRSV-MRV2 in Vero cells, a putative production cell line. Few nucleotide changes were observed on the level of viralpopulation consensus ; their biological significance remains to be established. The subconsensus population variability is alsobeing analysed, as minor variant of the virus quasispecies can affect the attenuation status of the live viral vaccines.

mumps virus, recombinant viral vaccines, reverse genetics technology

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