应用MAGE-1抗原肽体外诱导特异性细胞毒T淋巴细胞形成

Induction of cytotoxic Tlymphocytes from the peripheral blood of a hepatocellular carcinoma patient using a MAGE-1 peptide (NYKCRFPEI) in vitro

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。

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.