Induction of anti-carbohydrate antibodies by phage library-selected peptide mimics.
One of the prerequisites for the development of polysaccharide subunit vaccines is the induction of an efficient immune response to carbohydrate antigens like lipopolysaccharide (LPS) or capsular polysaccharide antigens of pathogens. In an attempt to overcome the problems that arise from the T-independent immune response induced by such antigens, selecting peptide sequences that mimic protective carbohydrate epitopes has been proposed. In this study, we investigate a new selection strategy for immunogenic peptide mimics using the phage-displayed peptide library technology. Two monoclonal antibodies (mAb) of the A isotype (mIgA), mIgA C5 and mIgA I3, specific for the O-antigen (O-Ag) part of the human pathogen Shigella flexneri serotype 5a LPS and protective against homologous infection were used to screen two phage-displayed nonapeptide libraries in pVIII. Using mIgA C5, 13 different specific clones were selected, and 6 using mIgA I3; 5 of the latter also interacted in enzyme-linked immunosorbent assay with the first mAb. All of the 19 clones selected were separately used to immunize mice, but only 2 of them, p100c (mIgA I3-specific) and p115 (interacting with both mIgA) were able to induce anti-O-Ag antibodies. The immune response was specific for the O-Ag of the S. flexneri serotype 5a, and also selectively recognized the corresponding bacterial strain. The amino acid sequences of p100c and p115 immunogenic peptide mimics were YKPLGALTH (flanked by two Cys residues) and KVPPWARTA, respectively. These results are the first example of immunogenic mimicry of carbohydrates by phage-displayed peptides, and indicate a new strategy of selection of immunogens for the development of anti-polysaccharide vaccines.
Phalipon
A
A
Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, Paris, France. phalipon@pasteur.fr
Folgori
A
A
Arondel
J
J
Sgaramella
G
G
Fortugno
P
P
Cortese
R
R
Sansonetti
P J
PJ
Felici
F
F
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006801:Humans; D007114:Immunization; D008070:Lipopolysaccharides / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D019081:O Antigens / Q000276:immunology*; D019151:Peptide Library; D010455:Peptides / Q000276:immunology*; D012763:Shigella flexneri / Q000276:immunology
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D008070:Lipopolysaccharides; D019081:O Antigens; D019151:Peptide Library; D010455:Peptides