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The Journal of Immunology, 2006, 176: 1686-1694.
Copyright © 2006 by The American Association of Immunologists

Characterization of Functional Oligosaccharide Mimics of the Shigella flexneri Serotype 2a O-Antigen: Implications for the Development of a Chemically Defined Glycoconjugate Vaccine1

Armelle Phalipon2,*, Corina Costachel*,{dagger}, Cyrille Grandjean{dagger}, Audrey Thuizat*, Catherine Guerreiro{dagger}, Myriam Tanguy*, Farida Nato{ddagger}, Brigitte Vulliez-Le Normand§, Frédéric Bélot{dagger}, Karen Wright{dagger}, Véronique Marcel-Peyre*,{dagger}, Philippe J. Sansonetti* and Laurence A. Mulard{dagger}

* Unité de Pathogénie Microbienne Moléculaire, Institut National de la Santé et de la Recherche Médicale Unité 389; {dagger} Unité de Chimie Organique, Centre National de la Recherche Scientifique Unité de Recherche Associée 2128; {ddagger} Plate-Forme 5, Production de Protéines Recombinantes et d’Anticorps; and § Unité d’Immunologie Structurale, Centre National de la Recherche Scientifique Unité de Recherche Associée 2185; Institut Pasteur, Paris, France

Protection against reinfection with noncapsulated Gram-negative bacteria, such as Shigella, an enteroinvasive bacterium responsible for bacillary dysentery, is mainly achieved by Abs specific for the O-Ag, the polysaccharide part of the LPS, the major bacterial surface Ag. The use of chemically defined glycoconjugates encompassing oligosaccharides mimicking the protective determinants carried by the O-Ag, thus expected to induce an efficient anti-LPS Ab response, has been considered an alternative to detoxified LPS-protein conjugate vaccines. The aim of this study was to identify such functional oligosaccharide mimics of the S. flexneri serotype 2a O-Ag. Using protective murine mAbs specific for S. flexneri serotype 2a and synthetic oligosaccharides designed to analyze the contribution of each sugar residue of the branched pentasaccharide repeating unit of the O-Ag, we demonstrated that the O-Ag exhibited an immunodominant serotype-specific determinant. We also showed that elongating the oligosaccharide sequence improved Ab recognition. From these antigenicity data, selected synthetic oligosaccharides were assessed for their potential to mimic the O-Ag by analyzing their immunogenicity in mice when coupled to tetanus toxoid via single point attachment. Our results demonstrated that induction of an efficient serotype 2a-specific anti-O-Ag Ab response was dependent on the length of the oligosaccharide sequence. A pentadecasaccharide representing three biological repeating units was identified as a potential candidate for further development of a chemically defined glycoconjugate vaccine against S. flexneri 2a infection.




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