摘要
目的:探讨肝癌(HCC)患者树突状细胞(DC)融合HCC细胞体外诱导同源T淋巴细胞产生特异性抗HCC免疫的效能. 方法:应用人重组粒细胞/巨噬细胞集落刺激因子(rhGM- CSF)和白介素-4(rhIL-4)对肝癌患者外周血单个核细胞进行体外诱导产生树突状细胞,流式细胞仪检测DC表面标志物表达水平,聚乙二醇融合DC与肝癌细胞HepG2,MTT法测定融合细胞(HepG2/DC)刺激同源T 淋巴细胞增生、分化能力,细胞毒性试验检测HepG2/ DC诱导的细胞毒T淋巴细胞(CTL)对HepG2的特异性杀伤作用. 结果:体外培养1 wk后的DC高度表达CDla,HLA- DR,CD54,CD80和CD86,融合细胞HepG2/DC刺激同源T淋巴细胞增值能力显著高于HepG2,HepG2+DC, DC及PBS,A值分别为0.816±0.019,0.541±0.020, 0.632±0.018,0.564±0.018,0.345±0.01 3(P<0.05), HepG2/DC活化的CTL对HepG2具有明显的特异性杀伤作用. 结论:HCC患者外周血DC融合HCC细胞可有效诱导同源T淋巴细胞产生特异性抗HCC免疫,可望成为一条HCC免疫治疗的有效途径.
AIM: To investigate the ability of fusions of HCC patient-derived dendritic cells (DC) with HCC cells to induce au-tologous T lymphocytes to elicit specific immunity against HCC in vitro. METHODS: Dendritic cells isolated from HCC patient peripheral blood were cultured and proliferated in vitro for one week by using recombinant human granulocyte/mac-rophage-colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4). Expression of DC surface markers was assessed by flow cytometry. Fusions of DC with HepG2 cells (HepG2/DC) were achieved by polythyleneglycol (PEG). The ability of HepG2/DC to stimulate proliferation and differentiation of autologous T lymphocytes was assessed by MTT method, and the specific killing efficacy of HepG2/DC-induced cytotoxic T lymphocytes (CTL) to HepG2 was evaluated. RESULTS: Following one week culture, DC presented a high-level expression of CD1a, HLA-DR, CD54, CD80 and CD86. Fusions had remarkably greater ability to stimulate proliferation of autologous T lymphocytes in comparison with HepG2, HepG2+DC, DC and PBS, with an A value of 0.816± 0.019 vs 0.541±0.020,0.632±0.018,0.564±0.018,0.345±0.013, respectively (P<0.05). The HepG2/DC-activated CTLs showed a potent specific killing efficacy to HepG2. CONCLUSION: Fusions of HCC patient-derived DC with HCC cells can effectively stimulate autologous T lymphocytes to elicit specific antitumor immunity against HCC, and may represent as a promising approach of immunotherapy for HCC.
出处
《世界华人消化杂志》
CAS
2004年第4期774-777,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金资助课题
No.30100180~~
