树突状细胞与卵巢癌细胞的融合及体外抗肿瘤作用的研究

目的 :比较脐血及以卵巢癌患者外周血来源的树突状细胞 (DC)的特点 ,研究DC 卵巢癌融合瘤苗的体外免疫应答效果。方法 :从脐血及卵巢癌患者外周血中诱导扩增DC ,从数量、形态、细胞表面标志及刺激增殖活性方面进行比较。体外培养卵巢癌患者癌细胞 ,将其与脐血DC在聚乙二醇 (PEG)介导下融合 ,活化自体T细胞 ,MTT法检测杀伤效果。结果 :体外诱导卵巢癌患者外周血DC ,每 2 0ml血能够获得 (0 .6～ 1.6 )× 10 6 个细胞 ;体外诱导脐血DC ,每 2 0ml血能够获得 (1.8～ 3.5 )× 10 6 个细胞 ,二者有明显差异 (P<0 .0 5 )。卵巢癌患者外周血DC在混和淋巴细胞反应 (mixedlymphocytereaction ,MLR)中显示了更为显著的刺激增殖活性。两者表达高水平的MHCII类分子和共刺激分子。经脐血DC 自体卵巢癌融合细胞活化的T细胞 ,对卵巢癌细胞株细胞的杀伤没有增加 ,而增强了对自体卵巢癌细胞的杀伤力。结论 :由脐血诱导可以获得更多数量的DC。脐血DC与自体卵巢癌细胞融合 ,能使肿瘤相关抗原有效提呈 ,激活T细胞 ,使它成为抗原特异性CTL ,有效杀伤自体癌细胞

Fusion of dendritic cells to ovarian carcinoma cells and the antitumor immunity study in vitro

Objective:To compare dendritic cells(DC) derived from cord blood a nd peripheral blood of patients with ovarian carcinoma.To study the anti-tumor immune response in vitro induced by fusion of DC to ovarian carcinoma cells.Methods:rhGM-CSF and rhIL-4 were used to generate DC in vitro from peripheral bl ood of patients with ovarian carcinoma.Meanwhile,rhGM-CSF and rhTNFα were use d to generate DC in vitro from cord blood.Comparison was made on population, m orphology, surface markers and stimulating ability between the two origins. Then DC derived from cord blood were fused to ovarian carcinoma cells, and the fusio n cells were cocultured with autologous PBMC in the presence of 200U/ml IL-2. A f ter 8 to 10 days, the cells were harvested and MTT test was carried out.R esults: The latter system yielded (1.8～3.5)×106 cells as the former one yielded (0. 6～1.6)×106 cells from 20ml blood. Yet DC derived from patient′s blood show ed m ore potent stimulating ability in MLR. Both expressed high levels of costimulato ry and MHCII molecules. Fusion of DC derived from cord blood to autologous ovar ian carcinoma cells activated T cells. And the anti-tumor ability was enhanced w hen autologous tumor cells were used as target cells.Conclusion:DC derived from cord blood can be a substitute of DC derived from patient′s blood in fusion to autol ogous ovarian carcinoma cells. And the fusions induce cytolytic T cell activity and lysis of autologous tumor cells by a MHC class I-restricted mechanism.