Naslov
Recombinant RSV and HCV vaccines based on mumps virus

Autori
Pali, Dorotea ; Košutić Gulija, Tanja ; Slović, Anamarija, Ivančić Jelečki, Jelena

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Knjiga sažetaka / - , 2021, 227-228

Skup
5. Simpozij studenata doktorskih studija PMF-a = 5th Faculty of Science PhD Student Symposium

Mjesto i datum
Zagreb, Hrvatska, 24.04.2021. - 25.04.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
mumps virus, reverse genetics technology

Sažetak
Attenuated mumps virus (Mumps orthorubulavirus, family Paramyxoviridae) is a promising candidate for the basis of recombinant vector vaccines because it elicits a sustained immune response and does not cause alterations in the host genome. Paramyxoviruses have single-stranded, nonsegmented, negative-sense RNA genome which is encapsidated by the nucleocapsid protein (NP) and further associated with RNA-dependant RNA polymerase (RdRp) consisting of phosphoprotein (P) and large protein (L). Transcription and replication of paramyxoviruses is mediated by the RdRp, which means that a successful production of virions in the host cell requires the presence of viral genome and simultaneous expression of proteins NP, P and L. For the production of mumps recombinant virus (vMRV2) and recombinant vector vaccines based on vMRV2 we are using a reverse genetics technology called rescue. This method is based on cotransfection of BSRT7 cells with four different plasmids: three expression plasmids that encode NP, P or L proteins and a plasmid with full-length genome of mumps vaccine strain L- Zagreb which does or does not include additional genes. We are using this technology to construct vector vaccines for hepatitis C virus (HCV) and human respiratory syncytial virus (RSV). For RSV vector vaccine, sequence encoding the ectodomain of RSV F protein fused to transmembrane and cytoplasmic region (TMD/CD) of mumps F protein was cloned into the genome of vMRV2. For HCV vaccine, we have inserted a) genomic region of HCV E1 and E2 proteins ; or b) sequences encoding ectodomains of HCV E1 and E2 proteins fused to TMD/CD region of mumps F protein. We expect that RSV and HCV genes will be expressed in infected cells and proteins with the addition of TMD/CD of mumps F protein will be incorporated in viral particles. By using rescue, we have produced recombinant viruses vMRV2 and vF RSV-MRV2 (vector vaccine for RSV) ; rescue for vCE1E2 HCV-MRV2 (vector vaccine for HCV, plasmid a) and vE1E2TMD HCV-MRV2 (plasmid b) is currently in progress. Viruses vMRV2 and vF RSV-MRV2 were propagated in Vero cell line, virus titer was determined and whole genome sequences were confirmed by next generation sequencing.

Izvorni jezik
Engleski

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