Generation of antibodies to human IL-12 and amphiregulin by immunization of Balb/c mice with diepitope multiple antigen peptides.
Six peptide sequences derived from the human proteins/oligopeptides IL-12, amphiregulin and FALL-39 were synthesized in order to raise specific antibodies in Balb/c mice. Although peptides are valuable tools for generating specific antibodies, they are often poor immunogens due to their small size and lack of relevant T-cell epitopes. To circumvent these limitations, the human peptides were co-synthesized in diepitope multiple antigen peptides (MAP) with a known H-2d-restricted T helper-cell epitope. The importance of including a T-cell epitope in the diepitope MAPs was demonstrated by the fact that only one of the human peptides was immunogenic as a monoepitope MAP, lacking the T-cell epitope. Conversely, all diepitope MAPs generated potent antibody responses to the desired human peptides as well as to the T-cell epitope. A certain degree of variability of the antibody responses to the diepitope MAPs indicated that the alterable component, i.e. the human B-cell epitope, influenced the T-cell help elicited by the T-cell epitope. Still, the relative conformity of the B-cell responses suggests that this strategy is generally applicable for a rational production of specific antibodies. Moreover, antiserum to four diepitope MAPs recognized the corresponding full-length human protein/oligopeptide as did monoclonal antibodies made against IL-12-and amphiregulin-based MAPs.
Ahlborg
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N
Department of Immunology, Stockholm University, Sweden.
Paulie
S
S
Braesch-Andersen
S
S
D000595:Amino Acid Sequence; D066258:Amphiregulin; D000818:Animals; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D023181:Antimicrobial Cationic Peptides; D066255:EGF Family of Proteins; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005260:Female; D006023:Glycoproteins / Q000138:chemical synthesis / Q000276:immunology*; D006133:Growth Substances / Q000138:chemical synthesis / Q000276:immunology*; D006801:Humans; D007114:Immunization; D036341:Intercellular Signaling Peptides and Proteins; D018664:Interleukin-12 / Q000138:chemical synthesis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology
C587848:AREG protein, human; D066258:Amphiregulin; D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D000941:Antigens; D023181:Antimicrobial Cationic Peptides; C587849:Areg protein, mouse; D066255:EGF Family of Proteins; D018984:Epitopes, T-Lymphocyte; C091425:FALL 39; D006023:Glycoproteins; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D018664:Interleukin-12