Two genes encoding human melanoma antigens MART-1 and gp100 recognized by HLA-A2 restricted melanoma reactive CTL derived from tumor infiltrating lymphocytes (TIL) were isolated by cDNA expression cloning methods. Multiple unmutated self peptides were identified as T cell epitopes in these melanocyte/melanoma specific proteins (2 from MART-1 and 5 from gp100). Most of these melanoma epitopes contain non-dominant anchor amino acids at the primary anchor positions and have intermediate binding affinity to HLA-A2.1. Melanoma reactive CTL were efficiently induced from PBL and TIL of patients by in vitro stimulation with PBMC pulsed with these epitopes. There is a significant correlation between vitiligo development and clinical response to IL2 based immunotherapy, suggesting that autoreactive T cells are involved in melanoma regression in vivo. These results have implications for understanding the nature of tumor antigens recognized by T cells and for the development of new cancer immunotherapies.