GENERATION OF SPECIFIC ANTIMELANOMA REACTIVITY BY STIMULATION OF HUMAN TUMOR-INFILTRATING LYMPHOCYTES WITH MAGE-1 SYNTHETIC PEPTIDE

The MAGE-1 gene encodes a tumor-specific antigen, MZ2-E, which is reco gnized by cloned, specific cytolytic T cells (CTL) derived from the pe ripheral blood of a patient with melanoma. We have produced a MAGE-1-s pecific CTL line derived from the tumor-infiltrating lymphocytes (TIL) of a melanoma patient by weekly restimulation with autologous EBV-B c ells pulsed with the synthetic HLA-A1-restricted MAGE-1 epitope nonape ptide EADPTGHSY. The 1277.A TIL line grew in long-term culture in low- dose interleukin-2 (IL-2) and IL-4, and exhibited antigen-specific, MH C-class-I-restricted lysis of HLA-A1-bearing MAGE-(1+) cell lines. Cyt olysis of target cells pulsed with the synthetic MAGE-1 decapeptide KE ADPTGHSY was superior to that of cells pulsed with the immunodominant nonapeptide. Single amino-acid or even side-chain substitutions in the immunodominant nonamer abrogated cytolysis. 1277.A TIL specifically s ecreted tumor necrosis factor a after co-incubation with HLA-A1-expres sing MAGE-(1+) cell lines or fresh tumor. These data suggest that tumo r-antigen-specific, MHC-restricted CTL may be grown from TIL in the pr esence of synthetic epitope peptides and expanded for adoptive immunot herapy in melanoma patients.