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目的探讨应用MAGE 1抗原肽治疗肝细胞肝癌 (HCC)的可行性。方法接受MAGE 1抗原九肽NYKCRFPEI孵育的外周血单个核细胞 (PBMC)经 30 0 0rad的射线照射后用作抗原呈递细胞 (APC) ,每隔 7d刺激HCC病人自体的PBMC 1次 ,进行混合淋巴细胞培养 (MLCs) ,共 4次后作为细胞毒T淋巴细胞 (CTL) ,应用乳酸脱氢酶 (LDH)释放分析法检测CTL对靶细胞的杀伤效应。结果从培养的第 1周至第 4周 ,淋巴细胞共增加至原细胞数的 32倍 ;在效应细胞∶靶细胞 (E∶T)为10∶1时 ,CTL对MAGE 1抗原九肽NYKCRFPEI孵育的自体淋巴母细胞的杀伤效应为 6 2 5 % ,对MAGE 1阳性、HLA A2 4阳性的HCC细胞株BEL740 5的杀伤效应为 40 2 % ,两者均明显高于对自体淋巴母细胞的杀伤效应 (17 9% )、和对MAGE 1阳性 ,HLA A2 4阴性的HCC细胞株HLE的杀伤效应(19 6 % ) ,及对MAGE 1阴性、HLA A2 4阴性的HCC细胞株QGY 770 1的杀伤效应 (1 6 % ) ;在E∶T为3 3∶1时 ,CTL对肽孵育的自体淋巴母细胞的杀伤效应为 5 3 6 % ,明显高于对自体淋巴母细胞的杀伤效应 (15 6 % )、对HLE的杀伤效应 (13% )和对QGY770 1的杀伤效应 (1% )。结论本实验结果表明 ,应用MAGE 1抗原肽NYKCRFPEI,能在体外从HCC病人的PBMC中有效地诱导出具有特异性杀伤能力的CTL。
Objective To investigate the feasibility of using MAGE 1 antigen peptide in the treatment of hepatocellular carcinoma (HCC). Methods Peripheral blood mononuclear cells (PBMCs) incubated with MAGE 1 nonapeptide NYKCRFPEI were exposed to 30 000 radions and used as antigen-presenting cells (APC). The autologous PBMC were stimulated once every 7 days with mixed lymphoid Cell culture (MLCs) were used as cytotoxic T lymphocytes (CTLs) four times. The killing effect of CTL on target cells was detected by lactate dehydrogenase (LDH) release assay. Results From the first week to the fourth week of culture, the number of lymphocytes increased to 32 times of that of the original cells. When the effector cell: target cell (E: T) was 10: 1, the CTLs were incubated with the MAGE1 nonapeptide NYKCRFPEI The lethal effect of autologous lymphoblasts was 65.5% and the killing effect on MAGE 1-positive, HLA-A2 4-positive HCC cell line BEL740 5 was 40.2%, both of which were significantly higher than that of autologous lymphoblasts (17 9%), killing effect (19 6%) on HLE cell line MAGE 1 positive and HLA A2 4 negative, and killing effect on MAGE 1 negative and HLA A2 4 negative HCC cell line QGY 770 1 (16%). When the ratio of E: T was 3 3:1, the killing effect of CTL on the autologous lymphoblastoid cells incubated with peptide was 536%, which was significantly higher than that of autologous lymphoblasts (15 6% ), Killing effect on HLE (13%) and killing effect on QGY770 1 (1%). Conclusion The results of this experiment show that the use of MAGE 1 antigen peptide NYKCRFPEI can effectively induce CTL with specific killing ability in PBMC of HCC patients in vitro.