人脐血树突状细胞抗神经胶质瘤的体外免疫效应研究

Antiglioma effect of dendritic cells derived from human cord blood in vitro

目的 研究肿瘤抗原致敏的树突状细胞 (DC)对神经胶质瘤细胞的免疫杀伤效应。方法 应用免疫磁珠分选脐血 CD34细胞 ,经 SCF+FL3+GM- CSF+IL- 4 +TNF- α的联合诱导培养 DC,采用相差显微镜观察树突状细胞的形态 ;流式细胞仪作 DC的表面标志检测 ;MTT比色法测定同种异型的混合淋巴细胞反应能力和诱导CTL 毒性的检测。结果 (1)脐血 CD34细胞在体外经细胞因子联合刺激后呈典型的 DC形态 ;(2 )流式检测 CD4 0、CD80和 CD86等成熟 DC特异性表面标志呈高表达 ,分别与诱导前比较 ,差异均有显著性 ;(3) MTT法测得脐血来源的 DC较外周血 DC刺激同种异体 T淋巴细胞增殖能力弱 ;经肿瘤抗原负载的 DC体外诱导出较强 CTL 毒性。结论 脐血 CD34细胞经体外扩增诱导的 DC具有典型的 CTL 毒性 ,为临床应用脐血来源

Objective To investigate the antiglioma e ffect of the CD34+ dendritic cells derived from human cord blood in vitro. Methods CD34+ mononuclear cells were isolated from neonatal cor d blood by ficoll centrifugation and magnetic se-paration technique, then cultu red with cytokines (SCF,FL-3,GM-CSF,IL-4,TNF-α). After two weeks of culture , the cells were checked by FACS. The allo-MLR and CTL were detected by MTT me thod.Results (1) CD34+ mononuclear cells from cord blood exh ibited distinctive morphological features of dendritic cell after culture. (2) F low cytometry demonstrated CD40, CD80, CD86 expressed on the mature DCs. (3)Allo -mixed lymphocyte reaction showed that DCs from cord blood stimulated by tumor antigens (SHG-44) induced stronger CTL reaction. Conclusion C D34 dendritic cells from cord blood are capable of inducing antiglioma respons e in vitro.