Joint production of prime/boost pairs of Fowlpox Virus and Modified Vaccinia Ankara recombinants carrying the same transgene

Elisa Soprana, Maddalena Panigada, Mathias Knauf, Antonia Radaelli, Luisa Vigevani, Alessio Palini, Chiara Villa, Mauro Malnati, Giulia Cassina, Reinhard Kurth, Stephen Norley, Antonio G. Siccardi

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)22-28
Number of pages7
JournalJournal of Virological Methods
Volume174
Issue number1-2
DOIs
Publication statusPublished - Jun 2011

Keywords

  • Prime/boost pairs
  • Recombinant poxviruses

ASJC Scopus subject areas

  • Virology

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