Although several MAGE-1 peptides have already been identified, the MAGE-1-encoded peptide presented by HLA-A24, which is the most common allele in Japanese population and is also frequently present in Caucasians, might have a wide applicability for immunotherapy using these peptides. To identify this potential peptide, we examined the induction of specific cytotoxic T lymphocytes (CTL) from the peripheral-blood mononuclear cells (PBMC) in HLA-A24 healthy donors by in vitro stimulation with MAGE-1-encoded synthetic peptides with a binding affinity for HLA-A24, by a simplified method. Of the 5 peptides tested, the highest HLA binder (NYKHCFPEI) was able to elicit CTL from unseparated PBMC by stimulation with freshly isolated, peptide-pulsed PMBC as antigen-presenting cells (APC) and by also using interleukin 7 and keyhole-limpet hemocyanin for a primary culture. The induced CTL could thus lyse HLA-A24 tumor cells expressing MAGE-1, as well as the peptide-pulsed target cells, in an HLA-class-I-restricted manner. By using the MAGE-1/HLA-A24 peptide, NYKHCFPEI, we found it possible to immunize many more patients, especially Japanese patients, by means of such peptide-based immunotherapeutic approaches to MAGE-1-positive malignant tumors.