Fabrication of influenza virus-like particles using M2 fusion proteins for imaging single viruses and designing vaccines

Vaccine. 2011 Sep 22;29(41):7163-72. doi: 10.1016/j.vaccine.2011.05.077. Epub 2011 Jun 7.

Abstract

Influenza virus-like particles (VLPs) are noninfectious and the assembly of influenza VLPs depends on the interactions of M1 proteins and/or other viral surface proteins, such as HA, NA, and M2, with the cellular lipid membranes. In this study we propose that M2 protein can be used as a molecular fabricator without disrupting the assembly of VLPs and while retaining the native structures of HA and NA envelope protein oligomers on the particle surfaces. First, we demonstrated that influenza VLPs can be fabricated by the M2 fusion of enhanced green fluorescent protein for imaging single virus entering A549 cells. Second, we engineered two molecular adjuvants (flagellin and profilin) fused to M2 protein to generate molecular adjuvanted VLPs. Theses molecular adjuvanted VLPs had stimulatory functions, including increasing TNF-α production and promoting the maturation of dendritic cells. Immunization of mice with molecular adjuvanted VLPs also enhanced the response of the neutralizing antibodies against homologous and heterologous H5N1 viruses. The results can provide useful information for imaging single viruses and designing novel vaccines against influenza virus infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / genetics
  • Adjuvants, Immunologic / metabolism
  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Viral / blood
  • Biotechnology / methods*
  • Cell Line
  • Dendritic Cells / immunology
  • Flagellin / genetics
  • Flagellin / metabolism
  • Genetic Engineering / methods
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hemagglutinin Glycoproteins, Influenza Virus / chemistry
  • Hemagglutinin Glycoproteins, Influenza Virus / genetics
  • Hemagglutinin Glycoproteins, Influenza Virus / immunology
  • Hemagglutinin Glycoproteins, Influenza Virus / metabolism
  • Humans
  • Influenza A Virus, H5N1 Subtype / genetics*
  • Influenza A Virus, H5N1 Subtype / immunology
  • Influenza A Virus, H5N1 Subtype / metabolism
  • Influenza Vaccines / administration & dosage
  • Influenza Vaccines / genetics*
  • Influenza Vaccines / immunology
  • Influenza Vaccines / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Neuraminidase / chemistry
  • Neuraminidase / genetics
  • Neuraminidase / immunology
  • Neuraminidase / metabolism
  • Orthomyxoviridae / genetics
  • Profilins / genetics
  • Profilins / metabolism
  • Protein Conformation
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Staining and Labeling
  • Technology, Pharmaceutical / methods*
  • Tumor Necrosis Factor-alpha / metabolism
  • Vaccines, Virosome / administration & dosage
  • Vaccines, Virosome / genetics
  • Vaccines, Virosome / immunology
  • Vaccines, Virosome / metabolism
  • Viral Matrix Proteins / genetics*
  • Viral Matrix Proteins / immunology
  • Viral Matrix Proteins / metabolism
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / immunology
  • Viral Proteins / metabolism

Substances

  • Adjuvants, Immunologic
  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Influenza Vaccines
  • M2 protein, Influenza A virus
  • Profilins
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • Vaccines, Virosome
  • Viral Matrix Proteins
  • Viral Proteins
  • enhanced green fluorescent protein
  • Flagellin
  • Green Fluorescent Proteins
  • NA protein, influenza A virus
  • Neuraminidase