title: Seamless replacement of Autographa californica multiple nucleopolyhedrovirus gp64 with each of five novel type II alphabaculovirus fusion sequences generates pseudotyped virus that fails to transduce mammalian cells.
abstract: Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a member of the type I alphabaculoviruses, is able to transduce and deliver a functional gene to a range of non-host cells, including many mammalian lines and primary cells, a property mediated by the envelope fusion protein GP64. AcMNPV is non-cytopathic and inherently replication deficient in non-host cells. As such, AcMNPV represents a possible new class of gene therapy vector with potential future clinical utility. Whilst not a problem for in vitro gene delivery, the broad tropism displayed for non-host cells is less desirable in a gene therapy vector. The fusion protein F of type II alphabaculoviruses can substitute functionally for GP64, and such pseudotyped viruses display a severely impaired capacity for non-host-cell transduction. Thus, surface decoration of such an F-pseudotyped AcMNPV with cell-binding ligands may restore transduction competence and generate vectors with desirable cell-targeting characteristics. By seamlessly swapping the native gp64 coding sequence with each of five sequences encoding different F proteins, a set of F-pseudotyped AcMNPV was generated. This report details their relative abilities both to functionally replace GP64 in viral growth and to transduce human Saos-2 and HeLa cells. All five supported viable infections in insect cell cultures and one, the Mamestra configurata NPV (MacoNPV) F pseudotype, could be amplified to titres close to those of native AcMNPV. In contrast, none was able to transduce the Saos-2 and HeLa cell lines. The robust support provided by MacoNPV F in virus production makes the corresponding pseudotype a viable scaffold to display surface ligands to direct selective mammalian cell targeting.
authors.affiliation: Pharmaceutical Science Research Division, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Marcel
authors.initials: M
authors.lastname: Westenberg
authors.affiliation: Pharmaceutical Science Research Division, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Helen M
authors.initials: HM
authors.lastname: Soedling
authors.affiliation: Pharmaceutical Science Research Division, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Nisha
authors.initials: N
authors.lastname: Hirani
authors.affiliation: Division of Cancer Studies, King's College London, Guy's Campus, London SE1 1UL, UK.
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Nicholson
authors.affiliation: Liver Research Group, Institute of Cellular Medicine, 4th Floor, Catherine Cookson Building Medical School, Newcastle University, Newcastle NE2 4HH, UK.
authors.forename: Derek A
authors.initials: DA
authors.lastname: Mann
authors.affiliation: Pharmaceutical Science Research Division, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Colin T
authors.initials: CT
authors.lastname: Dolphin
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D058507:Host Specificity; D006801:Humans; D007313:Insecta; D007915:Lepidoptera / Q000821:virology; D008322:Mammals; D011995:Recombination, Genetic; D014170:Transformation, Genetic; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins