摘要
目的将肝细胞癌(HCC)患者树突状细胞(DCs)与肝癌细胞HepG2融合制备瘤苗,检测其体外诱导同源T细胞产生特异性抗HepG2的免疫效应.方法以血细胞分离机分离富集HCC患者外周血单个核细胞(PBMCs),应用重组人粒细胞/巨噬细胞集落刺激因子(rhGM-CSF)、白细胞介素-4(rhlL-4)体外诱导培养DCs;聚乙二醇融合DCs与肝癌细胞HepG2,MTT法测定融合细胞(DCs/HepG2)刺激同源T淋巴细胞增殖分化能力,细胞毒性实验检测DCs/HepG2诱导细胞毒性T淋巴细胞(CTL)对HepG2的特异性杀伤作用.结果融合细胞DCs/HepG2高表达成熟DC表面分子,其中CD8390.4%,CD8087.7%,CD8684.4%,HLA-DR98.5%;其刺激同源T淋巴细胞增殖的能力显著高于HepG2和DCs;DCs/HepG2活化的CTL对HepG2具有显著杀伤作用,其杀伤率为(63.5±4.6)%(效靶比例为20∶1).结论HCC患者外周血DC融合HepG2细胞可有效诱导同源T淋巴细胞产生特异性抗HCC免疫效应,可能成为HCC免疫治疗的有效途径.
AIM： To investigate the fusion ability of hepatocellular carcinoma （HCC） patient-derived dendritic cells （DCs） with HCC cells （HepG2） in inducing autologous T lymphocytes to elicit specific immunity against HCC in vitro. METHODS： Peripheral blood mononuclear cells （PBMCs） from HCC patients were isolated by blood separator. In the presence of recombinant human granulocyte/macrophage-elone stimulating factor （rhGM-CSF） and interleukin-4 （rhIL-4）, PBMCs were cultured in vitro for 1 week to induce DCs. The expression of cell surface molecules was assessed by flow cytometry. The fusion cells of DCs with HepG2 cells （DCs/HepG2） were achieved by polyethylene glycol （PEG）. The ability of DCs/HepG2 to stimulate the proliferation of autologous T lymphocytes was evaluated by MTY assay and the specific lysis of HepG2 by DCs/HepG2 induced cytotoxic T lymphocytes （CTLs） was detected by cytotoxicity test. RESULTS： After fusion, DCs/HepG2 highly expressed surface molecules, ineluding CD83 90.4%, CD80 87.7%, CD86 84.4% and HLA- DR 98.5%, The fusion cells had a remarkably greater ability to stimulate the proliferation of autologous T lymphocytes in comparison with HepG2 and DCs. The DCs/HepG2-activated CTLs showed a potent specific lysis to HepG2 cells, which was （63.5 ± 4.6 ） % with an effector-target ratio 20 ： 1. CONCLUSION： The fusion of DCs derived from HCC patients with HepG2 cells can effectively stimulate autologous T lymphocytes to elicit specific antitumor immunity against HCC and may serve as a promising vaccine for immunotherapy of HCC.
出处
《第四军医大学学报》
北大核心
2006年第6期553-555,共3页
Journal of the Fourth Military Medical University
关键词
树突细胞
细胞融合
癌
肝细胞
免疫疗法
dendritic cells
cell fusion
carcinoma, hepatocellular
immunotherapy
