脐血树突状细胞介导的食管癌疫苗实验研究

目的　用杂交瘤技术制备脐血树突状细胞 (DC)和食管癌细胞融合疫苗 ,分析其生物学特征及体外诱导特异性免疫应答的能力。方法　 2 0 0 2 - 0 12 0 0 3- 0 3汕头大学医院第一附属医院分离脐血CD3 4 干细胞并诱导扩增为成熟DC ,并与EC10 9细胞融合 ,免疫磁珠法筛选EC10 9 DC ;流式细胞术鉴定疫苗表型 ;绘制疫苗生长曲线 ;MTT法测定瘤苗诱导淋巴细胞增殖能力及体外特异性免疫应答能力。结果　疫苗可体外生长 ,高表达CD80 、CD83 和CD86,并能有效刺激淋巴细胞增殖反应 ,体外诱导细胞毒T细胞 (CTL)对EC10 9的杀伤活性显著高于对照组 (P <0 0 5 )。结论　EC10 9 DC具备了刺激免疫细胞活化的分子表型 ,能在体外刺激淋巴细胞增殖和诱导特异性CTL杀伤作用

Experimental study of cancer vaccine prepared by fusions of esophageal carcinoma cell line with dendritic cell derived from cord blood

Objective To establish the cancer vaccine (EC109-DC) prepared by fusions of esophageal carcinoma cell line with dendritic cell derived from cord blood and study the biological characteristics and antitumor immunity.Methods CD 34 hematopoietic stem cells were isolated from cord blood using CD 34 Progenitor Cell Isolation Kit by magnetic cell sorting system (MACS);expended the cells as DC,and then fused with EC109 by PEG-3600;selected the fusion cells by MACS;observed the phenotypes and proliferation by flow cytometry and technique of cell culture in vitro. Lymphocytes proliferation reaction and the CTL cytotoxicity were examined with MTT.Results EC109-DC could proliferate slowly in vitro and highly expressed CD80,CD83 and CD86.Lymphocytes proliferation reaction and the specific cytotoxicity against EC109 induced by EC109-DC cells were significantly higher than the control groups (P<0.05).Conclusion EC109-DC vaccine acquires the immuno-stimulating phenotype and can significantly stimulate the lymphocytes proliferation reaction, and the CTL activity.