CLONING AND CHARACTERIZATION OF THE GENES ENCODING THE MURINE HOMOLOGS OF THE HUMAN-MELANOMA ANTIGENS MART1 AND GP100

The recent identification of genes encoding melanoma-associated antige ns has opened new possibilities for the development of cancer vaccines designed to cause the rejection of established tumors. To develop a s yngeneic animal model for evaluating antigen-specific vaccines in canc er therapy, the murine homologues of the human melanoma antigens MART1 and gp100, which were specifically recognized by tumor-infiltrating l ymphocytes from patients with melanoma, were cloned and sequenced from a murine B16 melanoma cDNA library. The open reading frames of murine MART1 and gp100 encode proteins of 113- and 626-amino acids with 68.8 and 77% identity to the respective human proteins. Comparison of the DNA sequences of the murine MART1 genes, derived from normal melanocyt es, the immortalized nontumorgenic melanocyte line Melan-a and the B16 melanoma, showed all to be identical. Northern and Western blot analy ses confirmed that both genes encoded products that were melanocyte li neage proteins. Mice immunized with murine MART1 or gp100 using recomb inant vaccinia virus failed to produce any detectable T-cell responses or protective immunity against B16 melanoma. In contrast, immunizatio n of mice with human gp100 using recombinant adenoviruses elicited T c ells specific for hgp100. but these T cells also cross reacted with B1 6 tumor in vitro and induced significant but weak protection against B 16 challenge. Immunization with human and mouse gp100 together [adenov irus type 2 (Ad2)-hgp100 plus recombinant vaccinia virus (rVV)-mgp100] , or immunization with human gp100 (Ad2-hgp100) and boosting with hete rologous vector (rVV-hgp100 or rVV-mgp100) or homologous vector (Ad2-h gp100), did not significantly enhance the protective response against B16 melanoma. These results may suggest that immunization with heterol ogous tumor antigen, rather than self, may be more effective as an imm unotherapeutic reagent in designing antigen-specific cancer vaccines.