| | | 肝癌细胞HepG2与肝癌患者树突状细胞融合瘤苗的体外效应
| | 药学本科毕业论文 作者： 张红梅，张利旺，刘文超，潘伯荣，斯晓明，任军
【关键词】 细胞融合
Effect of a vaccine prepared by fusion of HepG2 cells with dendritic cells from patients with hepatocellular carcinoma in vitro
【Abstract】 AIM: To investigate the fusion ability of hepatocellular carcinoma (HCC) patientderived 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/macrophageclone stimulating factor (rhGMCSF) and interleukin4 (rhIL4), 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 MTT 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, including CD83 90.4%, CD80 87.7%, CD86 84.4% and HLADR 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/HepG2activated CTLs showed a potent specific lysis to HepG2 cells, which was (63.5±4.6)% with an effectortarget 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.
【Keywords】 dendritic cells; cell fusion; carcinoma, hepatocellular; immunotherapy
【摘要】 目的药学本科毕业论文： 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的药学本科毕业论文免疫效应. 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(rhGMCSF)、白细胞介素4(rhlL4)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CD83 90.4%, CD80 87.7%, CD86 84.4%, HLADR 98.5%；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.
【关键词】 树突细胞；细胞融合；癌，肝细胞；免疫疗法
0引言
肝细胞癌（hepatocellular carcinoma, HCC）因缺乏特异性肿瘤抗原且抗原性弱，其生物治疗相对困难. 树突状细胞(dendritic cells, DCs)是体内功能最强的专职抗原提呈细胞，可摄取、加工处理和提呈抗原，并有效激活T细胞产生抗原特异性免疫应答［1］. 将DC与肝癌细胞融合，理论上可实现DC对肝癌细胞抗原最充分的提呈，有望获得令人满意的肝癌生物治疗效果. 我们利用细胞融合技术将DC与肝癌细胞融合，制备融合细胞疫苗，探讨其生物学特性及抗肿瘤活性，为肝癌DC疫苗临床应用提供实验基础及相关资料.
1材料和方法
1.1材料
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