中华医学杂志（英文版）2002，Vol.115Issue(7)：1002-1005,4.
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
Induction of cytotoxic T lymphocytes from the peripheral blood of a hepatocellular carcinoma patient using melanoma antigen-1 (MAGE-1) peptide
吕建锋 1冷希圣 1彭吉润 1牟东成 1庞学雯 2尚小英 2陈慰峰2
作者信息
- 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
- 折叠
摘要
Abstract
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 (HLAⅠ) 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 was 3.3∶1, the cytotoxicity of effector cells against the peptide pulsed autogenous lymphoblasts was 53.6%, which was much higher against autogenous lymphoblasts, HLE cells and QGY7701 cells at 15.6%, 13% and 1%, respectively.Conclusion The results demonstrate that cytotoxic T lymphocytes with the ability to specifically lyse target cells expressing both MAGE-1 and HLA-A24 could be successfully induced by the MAGE-1 peptide NYKCRFPEI in vitro. This indicates that a good result might be anticipated if this peptide is used as a tumor vaccine to treat HLA-A24 HCC patients.关键词
黑色素瘤抗原 细胞毒T淋巴细胞 肝细胞癌 免疫治疗Key words
MAGE-1/cytotoxic T lymphocytes/hepatocellular carcinoma/immunotherapy分类
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吕建锋,冷希圣,彭吉润,牟东成,庞学雯,尚小英,陈慰峰..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[J].中华医学杂志（英文版）,2002,115(7):1002-1005,4.
