Construction and characterization of insect cell-derived influenza VLP: cell binding, fusion, and EGFP incorporation

昆虫细胞来源的流感病毒样颗粒 (VLP) 的构建和表征:细胞结合、融合和 EGFP 掺入

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作者:Yi-Shin Pan, Hung-Ju Wei, Chung-Chieh Chang, Chung-Hung Lin, Ting-Shyang Wei, Suh-Chin Wu, Ding-Kwo Chang

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.

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