Construction and Characterization of Insect Cell-Derived Influenza VLP : Cell Binding, Fusion, and EGFP Incorporation

• Published in: BioMed research international Vol. 2010; no. 2010; pp. 1 - 11
• Main Authors: Pan, Yi-Shin, Wei, Hung-Ju, Chang, Chung-Chieh, Lin, Chung-Hung, Wei, Ting-Shyang, Wu, Suh-Chin, Chang, Ding-Kwo
• Format: Journal Article
• Language: English
• Published: Cairo, Egypt Hindawi Puplishing Corporation 01.01.2010; Hindawi Publishing Corporation; Dar al -Nasr -al-Llktruni; Hindawi Limited
• Subjects: Animals; Avian flu; Baculoviridae - genetics; Biological and medical sciences; Biomedical research; Cell Line; Cloning, Molecular; Endocytosis; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - metabolism; Human viral diseases; Humans; Infections; Infectious diseases; Jurkat Cells; Life sciences; Medical sciences; Microscopy, Fluorescence; Neuraminidase - chemistry; Neuraminidase - isolation & purification; Neuraminidase - metabolism; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Spodoptera - genetics; Surface Plasmon Resonance; Viral diseases; Viral diseases of the respiratory system and ent viral diseases; Viral Envelope Proteins - chemistry; Viral Envelope Proteins - isolation & purification; Viral Envelope Proteins - metabolism; Viral infections; Virion - chemistry; Virion - isolation & purification; Virion - metabolism; Virus Internalization; Infection; Characterization; Incorporation; Arthropoda; Viral disease; Influenza; Insecta; Invertebrata; Cell fusion; In vitro
• ISSN: 1110-7243; 2314-6133; 1110-7251; 2314-6141
• DOI: 10.1155/2010/506363

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.