title: GFP co-expression reduces the A33R gene expression driven by a fowlpox vector in replication permissive and non-permissive cell lines.
abstract: The development of an effective prophylactic vaccine is still necessary to improve the safety of the conventional although-discontinued smallpox vaccine, and to protect from the threat of deliberate release of variola virus. This need also arises from the number of new cases of animal orthopoxvirus infections each year, and to reduce the risk to animal handlers. Fowlpox (FP) recombinants only replicate in avian species and have been developed against human infectious diseases, as they can elicit an effective immune response, are not cross-reactive immunologically with vaccinia, and represent safer and more promising immunogens for immunocompromised individuals. The aim of this study was the characterisation of two new fowlpox recombinants expressing the A33R vaccinia virus gene either alone (FP(A33R)) or with the green fluorescent protein (FP(A33R-GFP)) to verify whether GFP can affect the expression of the transgene. The results show that both FP(A33R) and FP(A33R-GFP) can express A33R correctly, but A33R mRNA and protein synthesis are higher by FP(A33R) than by FP(A33R-GFP). Therefore, GFP co-expression does not prevent, but can reduce the level of a vaccine protein, and may affect the protective efficacy of the immune response.
authors.affiliation: Department of Medical Biotechnologies and Translational Medicine, University of Milan, via Vanvitelli 32, Milan, Italy. massimiliano.bissa@unimi.it
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bissa
authors.forename: Sole Maria
authors.initials: SM
authors.lastname: Pacchioni
authors.forename: Carlo
authors.initials: C
authors.lastname: Zanotto
authors.forename: Carlo
authors.initials: C
authors.lastname: De Giuli Morghen
authors.forename: Antonia
authors.initials: A
authors.lastname: Radaelli
mesh_terms: D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D005587:Fowlpox virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D018155:Orthopoxvirus / Q000276:immunology; D011213:Poxviridae Infections / Q000276:immunology / Q000517:prevention & control; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014616:Vaccinia virus / Q000276:immunology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*; D014779:Virus Replication
chemical_list: C446940:A33R protein, Vaccinia virus; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins