Expression of MAGE genes in osteosarcoma. Expression of MAGE genes that encode tumor-rejection antigens recognized by cytotoxic T lymphocytes with major histocompatibility complex class-I antigens was investigated in human osteosarcomas (20 cell lines and eight fresh tumor tissues). MAGE-1, 2, 3, 4, and 6 genes were expressed at the mRNA level in 11 (52.4%), 10 (47.6%), 10 (47.6%) one (4.8%), and 10 (47.6%) of 21 tumor cell lines, respectively, and in five (62.5%), six (75%), five (62.5%), one (12.5%), and five (62.5%) of eight fresh tumor tissues as determined by the reverse transcription-polymerase chain reaction method. MAGE-1 or 4 protein was detected by immunoblot analysis in eight of 11 or one of one tumor cell lines, respectively, where it was expressed at the mRNA level. Major histocompatibility complex class-I antigens were expressed in 19 of 21 tumor cell lines. These results suggest that MAGE tumor-rejection antigens are expressed in substantial numbers of osteosarcomas in a major histocompatibility class-I-restricted manner.