【摘要】 目的探讨获得大量乳腺癌患者自体树突状细胞(dendritic cells,DCs)和细胞因子诱导的杀伤细胞(cytokine-induced cells,CIK细胞)的可行性。方法实验组,乳腺癌患者经外周血干细胞动员,取50mL外周血分外周血单个核细胞(PBMC),用AIM-V无血清培养基培养DCs和CIK,并设健康人对照组,用RPMI1640完全营养培养基。结果实验组PBMC产出2×108个并诱导出1.2×107个DCs和2×109个CIK细胞,显著高于对照组(P<0.01)。DCs的CD86、CD11c和HLA-DR分子都有较高表达,CIK细胞CD3+、CD56+双阳性细胞达到14.41%。细胞毒实验提示CIK细胞有强大的杀瘤活性。结论乳腺癌患者经外周血干细胞动员,用AIM-V无血清系统培养,少量外周血可诱导大量优质自体DCs和CIK细胞。更多还原

【Abstract】 Objective To investigate the possibility of the generation of large numbers of autologous Dendritic cells and Cytokine-induced killer cells from breast cancer patients by peripheral blood stem cells mobilized. Methods Peripheral blood stem cells of breastcancer patients were mobilized by using rh GM-CSF. Peripheral blood mononuclear cells (PBMC) from 50 mL peripheral blood of breast cancer patients (experiment) and healthy donors (control) were isolated by Ficoll gradient centrifugation respectively. Dcs from adherent PBMC were generated by using rhGM-CSF、IL-4 and TNF-α in medium AIM-V (experiment) or RPMI 1640 (control). CIK cells from non-adherent PBMC were generated by using INF-γ, IL-1β, IL-2 and anti-CD3 MoAb in medium AIM-V (experiment) or RPMI 1 640 (control). Results The number of PBMC, DCs and CIK cells is 2×108, 1.2×107 and 2×109 in experimental group, which is significant higher than that in control group (P < 0.01). The percentage of CD3+CD56+ cells in CIK cells is 14.41%. DCs highly expressed CD86、CD11 cand HLA-DR in bothgroups. Cytotoxic activity of CIK cell against Bel-7402 is 20.36%, 26.62%, 34.09% and 36.12% in the rate of effect and target cells 5:1, 10:1, 20:1 and 40:1. Conclusion The study suggested that large numbers of full mature autologous Dendritic cells and Cytokine-induced killer cells can generate from breast cancer patients by peripheral blood stem cells mobilized and suit for adoptive immunotherapy of breast cancer patients.更多还原