The recent emergence and rapid spread of the pandemic H1N1 swine influenza virus reminded us once again of the need for a universal influenza vaccine that can elicit heterosubtypic protection. Here, we show the superior immunogenicity and immunoprotective capacity of the full-length matrix protein 2 ectodomain (M2e) peptide coupled to keyhole limpet haemocyanin (KLH) compared with the N-terminal 9 aa residues of M2e (SP1). Immunization with M2e–KLH protected mice against a lethal challenge with influenza A virus and significantly reduced weight loss and lung virus titres. In addition, passive transfer of serum raised in rabbits against M2e–KLH protected mice against a lethal influenza virus challenge, whereas serum from rabbits immunized with SP1–KLH did not. Nevertheless, immunofluorescence staining revealed that rabbit serum raised against SP1–KLH bound specifically to infected Madin–Darby canine kidney cells. We conclude that the peptide SP1 contains an immunogenic epitope that is not sufficient for immunoprotection.


Article metrics loading...

Loading full text...

Full text loading...



  1. Bucher, D., Popple, S., Baer, M., Mikhail, A., Gong, Y. F., Whitaker, C., Paoletti, E. & Judd, A.(1989). M protein (M1) of influenza virus: antigenic analysis and intracellular localization with monoclonal antibodies. J Virol 63, 3622–3633. [Google Scholar]
  2. Darnule, T. V., Likhite, V., Darnule, A. T., Turino, G. M. & Mandl, I.(1980). Enhancement of humoral immune response against human lung elastin peptides. Experientia 36, 606–607.[CrossRef] [Google Scholar]
  3. Dawood, F. S., Jain, S., Finelli, L., Shaw, M. W., Lindstrom, S., Garten, R. J., Gubareva, L. V., Xu, X., Bridges, C. B. & Uyeki, T. M.(2009). Emergence of a novel swine-origin influenza A (H1N1) virus in humans. N Engl J Med 360, 2605–2615.[CrossRef] [Google Scholar]
  4. De Filette, M., Min Jou, W., Birkett, A., Lyons, K., Schultz, B., Tonkyro, A., Resch, S. & Fiers, W.(2005). Universal influenza A vaccine: optimization of M2-based constructs. Virology 337, 149–161.[CrossRef] [Google Scholar]
  5. Fiore, A. E., Bridges, C. B. & Cox, N. J.(2009). Seasonal influenza vaccines. Curr Top Microbiol Immunol 333, 43–82. [Google Scholar]
  6. Grebe, K. M., Yewdell, J. W. & Bennink, J. R.(2008). Heterosubtypic immunity to influenza A virus: where do we stand? Microbes Infect 10, 1024–1029.[CrossRef] [Google Scholar]
  7. Hanly, W. C., Artwohl, J. E. & Bennett, B. T.(1995). Review of polyclonal antibody production procedures in mammals and poultry. ILAR J 37, 93–118.[CrossRef] [Google Scholar]
  8. Jegerlehner, A., Schmitz, N., Storni, T. & Bachmann, M. F.(2004). Influenza A vaccine based on the extracellular domain of M2: weak protection mediated via antibody-dependent NK cell activity. J Immunol 172, 5598–5605.[CrossRef] [Google Scholar]
  9. Lamb, R. A., Zebedee, S. L. & Richardson, C. D.(1985). Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface. Cell 40, 627–633.[CrossRef] [Google Scholar]
  10. Mera, K., Nagai, M., Brock, J. W., Fujiwara, Y., Murata, T., Maruyama, T., Baynes, J. W., Otagiri, M. & Nagai, R.(2008). Glutaraldehyde is an effective cross-linker for production of antibodies against advanced glycation end-products. J Immunol Methods 334, 82–90.[CrossRef] [Google Scholar]
  11. Mozdzanowska, K., Feng, J., Eid, M., Kragol, G., Cudic, M., Otvos, L., Jr & Gerhard, W.(2003). Induction of influenza type A virus-specific resistance by immunization of mice with a synthetic multiple antigenic peptide vaccine that contains ectodomains of matrix protein 2. Vaccine 21, 2616–2626.[CrossRef] [Google Scholar]
  12. Neirynck, S., Deroo, T., Saelens, X., Vanlandschoot, P., Jou, W. M. & Fiers, W.(1999). A universal influenza A vaccine based on the extracellular domain of the M2 protein. Nat Med 5, 1157–1163.[CrossRef] [Google Scholar]
  13. Pinto, L. H., Holsinger, L. J. & Lamb, R. A.(1992). Influenza virus M2 protein has ion channel activity. Cell 69, 517–528.[CrossRef] [Google Scholar]
  14. Reed, L. J. & Muench, H.(1938). A simple method for estimating fifty percent endpoints. Am J Hyg 27, 493–497. [Google Scholar]
  15. Schulze, I. T.(1970). The structure of influenza virus. I. The polypeptides of the virion. Virology 42, 890–904.[CrossRef] [Google Scholar]
  16. Wang, R., Song, A., Levin, J., Dennis, D., Zhang, N. J., Yoshida, H., Koriazova, L., Madura, L., Shapiro, L. & other authors(2008). Therapeutic potential of a fully human monoclonal antibody against influenza A virus M2 protein. Antiviral Res 80, 168–177.[CrossRef] [Google Scholar]
  17. Webster, R. G. & Hinshaw, V. S.(1977). Matrix protein from influenza A virus and its role in cross-protection in mice. Infect Immun 17, 561–566. [Google Scholar]
  18. Zebedee, S. L. & Lamb, R. A.(1989). Growth restriction of influenza A virus by M2 protein antibody is genetically linked to the M1 protein. Proc Natl Acad Sci U S A 86, 1061–1065.[CrossRef] [Google Scholar]

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error