脐血树突状细胞经卵巢癌细胞刺激后抗卵巢癌免疫应答的体外研究  

The anti-tumor immune response of impulsed cord dendritic cells induced by ovarian tumor cell-a in vitro research

作　　者：李琳[1] 张震宇[1] 王淑珍[1] 刘崇东[1] 翟妍[1] 刘晋伟[1] 

机构地区：[1]首都医科大学附属北京朝阳医院妇产科,100020

出　　处：《北京医学》2006年第8期452-454,共3页Beijing Medical Journal

基　　金：国家自然科学基金资助项目(编号39970770)

摘　　要：目的研究脐血来源的树突状细胞(DC)的体外扩增培养方法及诱导特异性抗卵巢癌细胞免疫效应。方法①从脐血中分离单个核细胞后,获得单核细胞(Mo)。以粒细胞-巨噬细胞集落剂(GM"CSF)、IL"4、集落刺激因子(SCF)和酪氨酸激酶受体III配体(Flt"3L)诱导分化扩增,共培养7d后用流式细胞仪进行表型分析;②诱导单核细胞分化的第3天加入人卵巢癌细胞株SKOV3的冻融抗原,共培养4d后获得负载肿瘤抗原的成熟DC;将致敏的DC与从脐血中分离的同种同体T淋巴细胞共培养6d,获得效应细胞;MTT法检测效应细胞对人卵巢癌细胞株SKOV3、人肝癌细胞株Hu"7的细胞毒作用;结果①脐血来源的Mo在细胞因子的作用下,7d后分化生成成熟的DC,高表达特异性抗原CD83、CD86、CD80、CD1a;②DC可负载并提呈肿瘤抗原,诱导肿瘤特异性细胞毒性T淋巴细胞(CTL)的产生,不同浓度的CTL对卵巢癌细胞株SKOV3均有特异性杀伤、抑制作用(P<0.05)。结论脐血中DC的前体细胞(即单核细胞)可在体外分化扩增为成熟的功能性DC,并可诱导出特异性杀伤卵巢癌细胞的免疫效应,是一种临床应用前景良好的、方便有效的肿瘤免疫治疗方法。Objective To investigate the capability of generation of dendritic cells （DC） from precursor cells of cord blood and inducing specific antitumor immune response. Methods ①The monocytes （Mo） were isolated from cord blood and cultured with GM-CSF,IL-4,SCF and Flt-3L （FL）. 7 days later, cell-surface phenotyping were examined by cytofluorimetry; ②The mature DCs with special tumor antigen （TAA） were collected on the 7th day. After culturing for 3 days, the mature DCs could activate allogeneic T cells to tumor specific cytotoxic T lymphocytes （CTL）. MTT assay was employed to detect the inhibition rate of CTL and its supematant on SKOV3 cell and Hu-7 cell, respectively. Results ①DCs precursors in cord blood could be induced to differentiate and mature in medium containing cytokines. After culturing for 7 days, the induced mature DCs had high expression of CD83,CD86,CD80 and CD1a; ②Mature DCs with TAA could activate native T cells to become tumor-specific CTL. Different cell amounts of CTL had significant inhibition or killing response or1 SKOV3 cells （P 〈 0.05）. Conclusions The Mo from cord blood can be induced to functional DCs which can activate native T lymphocyte to CTL and induce specific antitumor immune response. The functional DCs generated form cord blood have therapeutic potential in cellular therapy for ovarian cancer and may shed light in clinical tumor treatment.

关 键 词：树突状细胞 卵巢癌 免疫 

分 类 号：R737.31[医药卫生—肿瘤] R392.12[医药卫生—临床医学]