title: Construction and characterization of insect cell-derived influenza VLP: cell binding, fusion, and EGFP incorporation.
abstract: We have constructed virus-like particles (VLPs) harboring hemagglutinin (HA), neuraminidase (NA), matrix protein 1 (M1) ,and proton channel protein (M2) using baculovirus as a vector in the SF9 insect cell. The size of the expressed VLP was estimated to be ~100 nm by light scattering experiment and transmission electron microscopy. Recognition of HA on the VLP surface by the HA2-specific monoclonal antibody IIF4 at acidic pH, as probed by surface plasmon resonance, indicated the pH-induced structural rearrangement of HA. Uptake of the particle by A549 mediated by HA-sialylose receptor interaction was visualized by the fluorescent-labeled VLP. The HA-promoted cell-virus fusion activity was illustrated by fluorescence imaging on the Jurkat cells incubated with rhodamine-loaded VLP performed at fusogenic pH. Furthermore, the green fluorescence protein (GFP) was fused to NA to produce VLP with a pH-sensitive probe, expanding the use of VLP as an antigen carrier and a tool for viral tracking.
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors.forename: Yi-Shin
authors.initials: YS
authors.lastname: Pan
authors.forename: Hung-Ju
authors.initials: HJ
authors.lastname: Wei
authors.forename: Chung-Chieh
authors.initials: CC
authors.lastname: Chang
authors.forename: Chung-Hung
authors.initials: CH
authors.lastname: Lin
authors.forename: Ting-Shyang
authors.initials: TS
authors.lastname: Wei
authors.forename: Suh-Chin
authors.initials: SC
authors.lastname: Wu
authors.forename: Ding-Kwo
authors.initials: DK
authors.lastname: Chang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004705:Endocytosis; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D009439:Neuraminidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics; D020349:Surface Plasmon Resonance; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D014771:Virion / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D053586:Virus Internalization
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D009439:Neuraminidase