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

J Biomed Biotechnol. 2010:2010:506363. doi: 10.1155/2010/506363. Epub 2010 Dec 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cell Line
  • Cloning, Molecular
  • Endocytosis
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Jurkat Cells
  • Microscopy, Fluorescence
  • Neuraminidase / chemistry
  • Neuraminidase / isolation & purification
  • Neuraminidase / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Spodoptera / genetics
  • Surface Plasmon Resonance
  • Viral Envelope Proteins / chemistry*
  • Viral Envelope Proteins / isolation & purification
  • Viral Envelope Proteins / metabolism
  • Virion / chemistry*
  • Virion / isolation & purification
  • Virion / metabolism
  • Virus Internalization

Substances

  • Recombinant Proteins
  • Viral Envelope Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Neuraminidase