Strategies have been developed to characterize tumor antigens recognized by cytolytic T lymphocytes (CTL). We use a genetic approach based on the transfection of HLA genes and cDNA libraries in COS cells to isolate the gene producing the antigenic peptide. The tumor-specific expression of this gene can be evaluated by cDNA synthesis and quantitative PCR amplification. Transfection of fragments of the isolated gene allows the identification of the region encoding the antigenic peptide. Peptides are synthesized and tested for their ability to sensitize target cells to lysis by the CTL.