Generation of antibodies to human IL-12 and amphiregulin by immunization of Balb/c mice with diepitope multiple antigen peptides

Abstract

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.