title: Avian influenza virus hemagglutinin display on baculovirus envelope: cytoplasmic domain affects virus properties and vaccine potential.
abstract: Hemagglutinin (HA) is the major immunogen on the envelope of avian influenza virus (AIV). Therefore we constructed two recombinant baculoviruses: Bac-HA, expressing histidine-tagged HA with the cytoplasmic domain (CTD) derived from HA, and Bac-HA64, expressing histidine-tagged HA with the CTD derived from baculovirus envelope protein gp64. After infection, HA with either CTD was expressed and anchored on the plasma membrane of Sf-9 cells, as revealed by confocal microscopy. Immunogold electron microscopy demonstrated that both Bac-HA and Bac-HA64 displayed HA on the viral surface. However, analyses of purified viruses revealed that significantly more HA was incorporated into Bac-HA64 than into Bac-HA. In comparison with Bac-HA, Bac-HA64 significantly improved the gene delivery and transgene expression in mammalian cells, as determined by quantitative real-time polymerase chain reaction and flow cytometry. Bac-HA64 elicited significantly higher hemagglutination inhibition titers in mouse models than Bac-HA and the negative controls. These data collectively confirmed that the gp64 CTD, in comparison with HA CTD, resulted in more efficient HA incorporation into baculovirus, more efficient transgene delivery and expression, and elevated immunogenicity. This is the first report demonstrating the potential of HA-pseudotyped baculovirus as an avian influenza vaccine and that the choice of CTD tremendously affects baculovirus properties and vaccine efficacy.
authors: Department of Chemical Engineering, National Tsing Hua, University, Hsinchu, Taiwan.
authors: Ding-Gang
authors: DG
authors: Yang
authors: Yao-Chi
authors: YC
authors: Chung
authors: Yiu-Kay
authors: YK
authors: Lai
authors: Chia-Wei
authors: CW
authors: Lai
authors: Hung-Jen
authors: HJ
authors: Liu
authors: Yu-Chen
authors: YC
authors: Hu
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D001717:Birds / Q000821:virology; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006388:Hemagglutinins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000378:metabolism; D007252:Influenza Vaccines / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D016133:Polymerase Chain Reaction; D018411:Spodoptera; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D006388:Hemagglutinins; D007252:Influenza Vaccines; D014764:Viral Proteins; D049452:Green Fluorescent Proteins