ObjectiveTo use MAGE-1 antigen as tumor vaccine for the treatment of hepatocellular carcinoma (HCC).MethodsIrradiated peripheral blood mononuclear cells (PBMCs) pulsed with a MAGE-1 peptide (NYKCRFPEI) were used as antigen presenting cells (APC). The PBMCs from the same patient were stimulated with APCs on every 7th day for 4 times to elicit cytotoxic T lymphocytes (CTLs). The cytotoxicity of CTLs to various kinds of target cells was detected with the method of lactate dehydrogenase (LDH) releasing assay. Results The number of PBMCs increased by 32 folds during 28 days of culture. When effective cells: target cells (E∶T) was 10∶1, CTLs exhibited 62.5% cytotoxicity against autogenous lymphoblasts pulsed with the peptide of MAGE-1 antigen (NYKCRFPEI), 40.25% cytotoxicity against cells of BEL7405, a HCC cell line expressing both MAGE-1 and HLA-A24, compared with 17.88% cytolysis observed against autogenous lymphoblasts, 19.55% against cells of HLE, a HCC cell line expressing MAGE-1, negatively expressing HLA-A24. The cytotoxicity against cells of QGY7701, a HCC cell line expressing neither MAGE-1 nor HLA-A24 was much lower (1.6%). When E∶T was 3.3∶1, the cytotoxicity of CTL against peptide pulsed lymphoblasts was 53.6%, while against autogenous lymphoblasts, cells of HLE and cells of QGY7701 was much lower, 15.6%, 13% and 1% respectively. After 4h culture, most cells of BEL7405 were adhered with several CTL, few cells of QGY7701 were adhere with CTL. ConclusionsMAGE-1 peptide NYKCRFPEI in vitro successfully induced CTL with the ability of specifically killing target cells expressing both MAGE-1 and HLA-A24.