The specificity of antibodies raised against a T cell peptide is influenced by peptide amidation.
Peptides used for immunization are designed on the basis of combination of B and T cell epitopes. They are sometimes acetylated and amidated in order to mimic the protein insertion of the B cell epitope, but to our knowledge the effect of modifying the N- and C-termini is not clearly identified. In this paper, we have investigated in detail the influence of amidation and acetylation on the immunogenic properties of the T cell epitope 24-36 which is derived from a snake neurotoxin. Acetylation enhanced the capacity of the peptides to bind to I-Ed and to stimulate specific T cells in vitro but both modifications did not influence in vivo the T cell priming ability of the peptides. However, amidation of the peptides 24-36 provoked a dramatic effect on the antibody specificity they elicited, whereas acetylation did not. Antibodies recruited by amidated peptides weakly recognized the non amidated ones, while the latter elicited antibodies which hardly bind to the former. These results show how a subtle chemical change of a peptide immunogen modifies the reactivity of the elicited antibodies in an unrelated manner from the peptide MHC II binding ability and T cell stimulating capacity. We thus amplify the previously described polarity of chimeric TB peptides that raise antibodies mainly against their C-terminal part. Finally, these results may also facilitate the choice of the status of N and C termini of the peptides designed for immunization which at present have their extremities indifferently free or modified by acetylation and/or amidation.
Maillère
B
B
CEA, Département d'Ingénierie et d'Etudes des Protéines, CEA-Saclay, Gif-sur-Yvette, France. maillere@cea.fr
Hervé
M
M
D000107:Acetylation; D000577:Amides / Q000378:metabolism; D000818:Animals; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
D000577:Amides; D018984:Epitopes, T-Lymphocyte; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D010455:Peptides