Peptide mimics of a conserved H5N1 avian influenza virus neutralization site

Biochem J. 2009 Apr 1;419(1):133-9. doi: 10.1042/BJ20080083.

Abstract

A panel of 52 murine monoclonal antibodies was found to recognize antigenic determinants that had been conserved among all major genetic subgroups of the H5N1 avian influenza virus prevalent since 1997. We screened a phage display library for peptides recognized by one such antibody (8H5). We analysed the specificity of 8H5 for reactive peptides presented as fusion proteins of HBc (hepatitis B core protein) and HEV (hepatitis E virus) structural protein, p239. This was then related to the specificity of the native HA (haemagglutinin) molecule by virtue of the capacity of fusion proteins to compete for 8H5 binding with different strains of H5N1 virus and the reactivity of antisera generated against fusion proteins to bind native HA molecules, and to inhibit haemagglutination and arrest infection by the virus. Nine reactive peptides of different amino acid sequences were identified, six of which were also reactive with the antibody in association with HBc and four were in association with p239. Binding occurred with the dimeric form of the four p239-fusion proteins and one of the HBc-fusion proteins, but not with the monomeric form. The HBc-fusion proteins blocked 8H5 binding with four strains of H5N1 influenza virus. Mouse antisera generated against fusion proteins bound to HA molecules, but did not inhibit haemagglutination or arrest H5N1 infection. Our findings indicate that 8H5 recognizes discontinuous sites presented by secondary and possibly higher structural orders of the peptides in spatially favourable positions for binding with the antibody, and that the peptides partially mimic the native 8H5 epitopes on the H5N1 virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Blotting, Western
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Hemagglutination
  • Hepatitis B Core Antigens / genetics
  • Hepatitis B Core Antigens / immunology
  • Hepatitis B Core Antigens / metabolism
  • Hepatitis Viruses / genetics
  • Hepatitis Viruses / immunology
  • Hepatitis Viruses / metabolism
  • Immune Sera / immunology
  • Influenza A Virus, H5N1 Subtype / genetics
  • Influenza A Virus, H5N1 Subtype / immunology*
  • Influenza A Virus, H5N1 Subtype / metabolism*
  • Peptide Library
  • Peptides / genetics
  • Peptides / immunology*
  • Peptides / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Spodoptera
  • Viral Proteins / genetics
  • Viral Proteins / immunology
  • Viral Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Hepatitis B Core Antigens
  • Immune Sera
  • Peptide Library
  • Peptides
  • Recombinant Fusion Proteins
  • Viral Proteins