Induction of cytotoxic T lymphocytes from the peripheral blood of a hepatocellular carcinoma patient using melanoma antigen-1 (MAGE-1) peptide  

应用MAGE- 1抗原肽从肝癌病人外周血中诱导产生特异性细胞毒T淋巴细胞(英文)

作　　者：吕建锋[1] 冷希圣[1] 彭吉润[1] 牟东成[1] 庞学雯[2] 商小英[2] 陈慰峰[2] 

机构地区：[1]北京大学人民医院肝胆外科,北京100044 [2]北京大学医学部免疫学系,北京100044

出　　处：《Chinese Medical Journal》2002年第7期1002-1005,145-146,共4页中华医学杂志（英文版）

摘　　要：OBJECTIVE: To investigate the possibility of using melanoma antigen-1 (MAGE-1) peptide as a tumor vaccine to treat hepatocellular carcinoma (HCC). METHODS: The expressions of MAGE-1 in 8 HCC cell lines and in liver cancer tissue from a patient were detected using RT-PCR. The type of human leucocyte antigen I(HLA I) of both 8 HCC cell lines and peripheral blood mononuclear cells of the patient was detected using a microcytotoxicity method to screen out target cell lines for the cytotoxicity assay. Peripheral blood mononuclear cells from the HCC patient pulsed with an MAGE-1 peptide (NYKCRFPEI) were used as antigen presenting cells. Autogenous peripheral blood mononuclear cells were stimulated with antigen presenting cells every 7 days for 4 times to elicit cytotoxic T lymphocytes. The phenotype of effector cells was analyzed using flow cytometry. The cytotoxicity of effector cells was detected with a lactate dehydrogenase releasing assay. RESULTS: The expressions of both MAGE-1 and HLA-A24 were detected in BEL7405 cell line which were used as the positive target cell line in the cytotoxicity assay. The expression of MAGE-1 alone was detected in HLE, BEL7402, BEL7404, QGY7703 and SMMC7721 cell lines, and the expression of neither MAGE-1 nor HLA-A24 was shown in QGY 7701 and HpG2 cell lines. The last 7 cell lines could be used as negative target cell lines in the cytotoxicity assay. Peripheral blood mononuclear cells expanded 32 folds during 28-day culture. The ratio of CD3(+) T cells increased by 16% (from 54% to 70%), and the ratio of CD8(+) T cells increased by 20% (from 36% to 56%) during 28-day culture. When the ratio of effector cells to target cells was 10:1, effector cells exhibited 62.5% cytotoxicity against autogenous lymphoblasts pulsed with the peptide (NYKCRFPEI) of MAGE-1 antigen, 40.25% cytotoxicity against BEL7405 cells, compared with 17.88% cytolysis observed against autogenous lymphoblasts, 19.55% against HLE cells, and 1.6% against QGY7701 cells. When the ratio of effector cells to target cells wa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　应用RT PCR法检测MAGE 1在 8种HCC细胞株中的表达 ,筛选杀伤实验的靶细胞 ;将MAGE 1抗原肽NYKCRFPEI孵育的外周血单个核白细胞经照射后作为抗原呈递细胞,每隔7天刺激HCC病人自体外周血单个核白细胞一次,共4次后作为效应T淋巴细胞,应用流式细胞仪检测培养前后淋巴细胞的表型变化,应用乳酸脱氢酶法检测效应T细胞对靶细胞的杀伤效应。结果;;HCC细胞株BEL7405MAGE1和HLAA24均阳性表达,可用作杀伤实验的阳性靶细胞,HLE等其它7种细胞不能双表达,可用作杀伤实验的阴性对照细胞;;培养28天,淋巴细胞增加31倍;;培养28天,CD3+T和CD8+T分别增加16%和20%;;在效∶靶为10∶1时,效应T细胞对MAGE1抗原九肽NYKCRFPEI孵育的自体淋巴母细胞、BEL7405的杀伤效应分别为625%和4025%,均明显高于对自体淋巴母细胞(1788%)、HLE(1955%)及QGY7701的杀伤效应(16%);;在效∶靶为33∶1时,效应T细胞对肽孵育的自体淋巴母细胞的杀伤效应为536%,明显高于对自体淋巴母细胞(156%)、HLE(13%)和QGY7701的杀伤效应(1%)。结论;;本实验结果表明,应用MAGE1抗原肽NYKCRFPEI,在体外从HCC病人的外周血单个核白细胞中成功地诱导出具有特异性杀伤能力的效应T细胞,为应用MAGE1抗原肽对HCC?

关 键 词：Adult Cancer Vaccines Carcinoma  Hepatocellular HLA-A Antigens Humans Liver Neoplasms Male Neoplasm Proteins RNA  Messenger T-Lymphocytes  Cytotoxic Tumor Cells  Cultured 

分 类 号：R735.7[医药卫生—肿瘤]