title: Joint production of prime/boost pairs of Fowlpox Virus and Modified Vaccinia Ankara recombinants carrying the same transgene.
abstract: Pairs of recombinant MVA (Modified Vaccinia Ankara) and FPV (Fowlpox Virus) expressing the same transgene are reasonable candidates for prime/boost regimens, because cross-reacting immune responses between the two vectors, both non-replicative in mammalian hosts, are very limited. The acceptor virus FPD-Red, a derivative of FPV, carrying a red fluorescent protein gene flanked by the homology regions of MVA deletion III, was constructed. The same MVA Transfer Plasmid Green, designed to insert transgenes into the MVA deletion III locus, can therefore be used to transfer transgenes into both acceptor viruses MVA-Red and FPD-Red with the described recently Red-to-Green gene swapping method. Cells infected by either recombinant virus can be sorted differentially by a simple and reliable FACS-based purification protocol. The procedure is carried out in primary chick embryo fibroblasts grown in serum-free media and was applied to the production of three rMVA/rFPV pairs expressing the H5N1 avian influenza antigens M1, M2 and NP. The viral genes were human codon-optimized and expressed at high levels in both chick and mammalian cells. Both single-step and multiple-step growth analyses showed no significant differences in growth due to the transgenes in either rMVA or rFPV derivatives.
authors.affiliation: San Raffaele Scientific Institute, Milano, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Soprana
authors.forename: Maddalena
authors.initials: M
authors.lastname: Panigada
authors.forename: Mathias
authors.initials: M
authors.lastname: Knauf
authors.forename: Antonia
authors.initials: A
authors.lastname: Radaelli
authors.forename: Luisa
authors.initials: L
authors.lastname: Vigevani
authors.forename: Alessio
authors.initials: A
authors.lastname: Palini
authors.forename: Chiara
authors.initials: C
authors.lastname: Villa
authors.forename: Mauro
authors.initials: M
authors.lastname: Malnati
authors.forename: Giulia
authors.initials: G
authors.lastname: Cassina
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kurth
authors.forename: Stephen
authors.initials: S
authors.lastname: Norley
authors.forename: Antonio G
authors.initials: AG
authors.lastname: Siccardi
mesh_terms: D000818:Animals; D002460:Cell Line; D002645:Chickens; D004337:Drug Carriers; D005587:Fowlpox virus / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes; D014614:Vaccines, Synthetic / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics*
chemical_list: D004337:Drug Carriers; D014614:Vaccines, Synthetic; D014764:Viral Proteins; D014765:Viral Vaccines