title: Paramyxovirus-based production of Rift Valley fever virus replicon particles.
abstract: Replicon-particle-based vaccines combine the efficacy of live-attenuated vaccines with the safety of inactivated or subunit vaccines. Recently, we developed Rift Valley fever virus (RVFV) replicon particles, also known as nonspreading RVFV (NSR), and demonstrated that a single vaccination with these particles can confer sterile immunity in target animals. NSR particles can be produced by transfection of replicon cells, which stably maintain replicating RVFV S and L genome segments, with an expression plasmid encoding the RVFV glycoproteins, Gn and Gc, normally encoded by the M-genome segment. Here, we explored the possibility to produce NSR with the use of a helper virus. We show that replicon cells infected with a Newcastle disease virus expressing Gn and Gc (NDV-GnGc) were able to produce high levels of NSR particles. In addition, using reverse genetics and site-directed mutagenesis, we were able to create an NDV-GnGc variant that lacks the NDV fusion protein and contains two amino acid substitutions in, respectively, Gn and HN. The resulting virus uses a unique entry pathway that facilitates the efficient production of NSR in a one-component system. The novel system provides a promising alternative for transfection-based NSR production.
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Wichgers Schreur
authors.affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, the Netherlands.|Department of Virology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Nadia
authors.initials: N
authors.lastname: Oreshkova
authors.affiliation: Department of Infection Biology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Frank
authors.initials: F
authors.lastname: Harders
authors.affiliation: Department of Infection Biology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Alex
authors.initials: A
authors.lastname: Bossers
authors.affiliation: Department of Infectious Diseases and Immunology, Virology Division, Faculty of Veterinary Medicine, Utrecht University, Utrecht, the Netherlands.|Department of Virology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Rob J M
authors.initials: RJM
authors.lastname: Moormann
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen University and Research Centre, Lelystad, the Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kortekaas
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009522:Newcastle disease virus / Q000276:immunology*; D012295:Rift Valley Fever / Q000517:prevention & control*; D012296:Rift Valley fever virus / Q000502:physiology*; D014765:Viral Vaccines / Q000276:immunology*; D014779:Virus Replication
chemical_list: D014765:Viral Vaccines