医学论文-大量培养诱导乳腺癌患者自体 DC 和 CIK 细胞的研究.docVIP

医学论文-大量培养诱导乳腺癌患者自体 DC 和 CIK 细胞的研究 ???????????????????????? 作者：王先明 何劲松 陈伟财 王敏 闵军 邵静 【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 50 mL 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 RPMI 1640 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。 【关键词】? 乳腺癌；干细胞动员；树突状细胞；细胞因子诱导的杀伤细胞 ??? 〔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? generatedby? using? rhGM-CSF、IL-4? and? TNF-α? in? medium? AIM-V? (experiment)? or? RPMI? 1 640? (control).? CIK? cells? fromnon-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? both groups.? Cytotoxic? activity? of? CIK? cell? against? Bel-7 402? 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? sugg