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J. Biol. Chem., Vol. 283, Issue 17, 11382-11387, April 25, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/17/11382    most recent M800650200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by De Filette, M. Articles by Saelens, X. Search for Related Content PubMed PubMed Citation Articles by De Filette, M. Articles by Saelens, X. Social Bookmarking                      What's this?

An Influenza A Vaccine Based on Tetrameric Ectodomain of Matrix Protein 2^*

Marina De Filette, Wouter Martens, Kenny Roose, Tom Deroo, Frederik Vervalle, Mostafa Bentahir, Joel Vandekerckhove^¶||, Walter Fiers, and Xavier Saelens¹

From the Departments of Molecular Biomedical Research and ^¶Medical Protein Research, Vlaams Instituut voor Biotechnologie (VIB), B9052 Ghent, Belgium and the Departments of Molecular Biology and ^||Biochemistry, Ghent University, B9052 Ghent, Belgium

Matrix protein 2 (M2) of influenza A is a tetrameric type III membrane protein that functions as a proton-selective channel. The extracellular domain (M2e) has remained nearly invariable since the first human influenza strain was isolated in 1933. By linking a modified form of the leucine zipper of the yeast transcription factor GCN4 to M2e, we obtained a recombinant tetrameric protein, M2e-tGCN4. This protein mimics the quaternary structure of the ectodomain of the natural M2 protein. M2e-tGCN4 was purified, biochemically characterized, and used to immunize BALB/c mice. High M2e-specific serum IgG antibody titers were obtained following either intraperitoneal or intranasal administration. Immunized mice were protected fully against a potentially lethal influenza A virus challenge. Antibodies raised by M2e-tGCN4 immunization specifically bound to the surface of influenza-infected cells and to an M2-expressing cell line. Using a M2e peptide competition enzyme-linked immunosorbent assay with M2-expressing cells as target, we obtained evidence that M2e-tGCN4 induces antibodies that are specific for the native tetrameric M2 ectodomain. Therefore, fusion of an oligomerization domain to the extracellular part of a transmembrane protein allows it to mimic the natural quaternary structure and can promote the induction of oligomer-specific antibodies.

Received for publication, January 24, 2008

^* This work was supported by National Institutes of Health Grant 5R01AI055632. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S4.

¹ To whom correspondence should be addressed: Dept. of Molecular Biomedical Research, VIB and Ghent University, Technologiepark 927, B9052 Ghent, Belgium. Tel.: 32-9-33-13-620; Fax: 32-9-33-13-609; E-mail: xavier.saelens{at}dmbr.ugent.be.

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