Abstract
BackgroundInfluenza viruses are characterized by their highly variable surface proteins HA and NA. The third surface protein M2 is a nearly invariant protein in all Influenza A strains. Despite extensive studies in other animal models, this study is the first to describe the use of recombinant M2 protein and a peptide coding for the extracellular part of the M2 protein (M2e) to vaccinate poultry.MethodsFour groups of layer chickens received a prime-boost vaccination with recombinant M2 protein, M2e, a tetrameric construct from M2e peptide bound to streptavidin and a control tetrameric construct formulated with Stimune adjuvant.ResultsWe determined the M2-specific antibody (Ab) responses in the serum before vaccination, three weeks after vaccination and two weeks after booster, at days 21, 42 and 56 of age. The group vaccinated with the M2 protein in combination with Stimune adjuvant showed a significant Ab response to the complete M2 protein as compared to the other groups. In addition an increased Ab response to M2e peptide was found in the group vaccinated with the M2e tetrameric construct. None of the vaccinated animals showed seroconversion to AI in a commercial ELISA. Finally no Ab’s were found that bound to M2 expressed on in vitro AI infected MDCK cells.ConclusionAlthough Ab’s are formed against the M2 protein and to Streptavidin bound M2e peptide in a tetrameric conformation these Ab’s do not recognize of M2 on the virus or on infected cells.
Highlights
Influenza viruses are characterized by their highly variable surface proteins HA and NA
Wu et al found that M2 protein (M2e) alone was a poor immunogen which did not elicit a significant immune response in mice, while combined with aluminim or Freund adjuvant the peptide was immunogenic and vaccination protected against lethal dose of influenza virus challenge [11]
We report a study in which we vaccinated chickens with Stimune adjuvanted full length matrix 2 (M2) protein, M2e peptide and Streptavidin bound M2e peptide in a tetrameric conformation
Summary
Influenza viruses are characterized by their highly variable surface proteins HA and NA. Wu et al found that M2e alone was a poor immunogen which did not elicit a significant immune response in mice, while combined with aluminim or Freund adjuvant the peptide was immunogenic and vaccination protected against lethal dose of influenza virus challenge [11]. In chickens Nayak et al tested recombinant NDV vectors that expressed each of the three surface proteins of high pathogenic AIV. They found no indication for M2 to be immunogenic or protective [21]. Zhang et al used a plasmid coding for an M2 protein of which the transmembrane region was deleted They found that chicken C3d enhanced the humoral immunity against AIV M2 protein, be it with a poor protection ratio [22]. Layton et al (2009) showed that Salmonella vectored vaccines expressing a M2e epitope in association with CD154 are effective at protecting chickens against LPAI, but not against HPAI [23]
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