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 speci fic antibodies in Balb/c mice. Although peptides are valuable tools fo r generating specific antibodies, they are often poor immunogens due t o their small size and lack of relevant T-cell epitopes. To circumvent these limitations, the human peptides were co-synthesized in diepitop e multiple antigen peptides (MAP) with a known H-2(d)-restricted T hel per-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 epi tope. Conversely, all diepitope MAPs generated potent antibody respons es to the desired human peptides as well as to the T-cell epitope. A c ertain degree of variability of the antibody responses to the diepitop e MAPs indicated that the alterable component, i.e. the human B-cell e pitope, influenced the T-cell help elicited by the T-cell epitope. Sti ll, the relative conformity of the B-cell responses suggests that this strategy is generally applicable for a rational production of specifi c antibodies. Moreover, antiserum to four diepitope MAPs recognized th e corresponding full-length human protein/oligopeptides did monoclonal antibodies made against n-12- and amphiregulin-based MAPs.