卵巢癌细胞和脐血树突状细胞融合瘤苗的体外培养与免疫原性的研究  

The Anti-tumor Immunity and Cultivation of the Fused Cells of Ovarian Cancer and Dendritic Cells Derived from Cord Blood——An In-vitro Study

作　　者：李金凤[1] 张震宇[1] 王淑珍[1] 刘晋玮[1] 

机构地区：[1]首都医科大学附属北京朝阳医院妇产科

出　　处：《首都医科大学学报》2007年第5期604-608,共5页Journal of Capital Medical University

基　　金：国家自然科学基金(39970770)资助项目~~

摘　　要：目的将脐血来源的树突状细胞(DC)与人卵巢癌细胞株SKOV3细胞相融合,获得SKOV3/DC融合细胞,分析其生物学特性及其免疫原性。方法①利用PKH26红色荧光染料标记脐血来源的DC,用聚乙二醇法(PEG)融合DC与人卵巢癌细胞株SKOV3细胞,用流式细胞仪分选融合细胞(SKOV3/DC)。②应用细胞培养技术、流式细胞术及光学显微镜检测SKOV3/DC的生长特性和形态学特征,并分析其免疫原性。结果①脐血来源的单核细胞(Mo)在相关细胞因子的作用下,可诱导分化为DC,其可表达表面抗原CD1a、CD80、CD86、HLA-DR及MHC-Ⅰ,不表达CA125。②人卵巢癌细胞株SKOV3细胞为CA125及MHC-Ⅰ双阳性细胞,不表达CD1a、CD80、CD86和HLA-DR分子。③树突状细胞和SKOV3按10∶1比例融合,融合率约为8.5%。SKOV3/DC形态与亲本肿瘤细胞相似,CA125抗原表达呈阳性,并且高表达CD1a、CD80、CD86、HLA-DR和MHC-Ⅰ分子。SKOV3/DC在体外能分裂增殖,但增殖活性明显低于SKOV3细胞。结论①脐血中的DC前体细胞(单核细胞)可在体外分化扩增为成熟的功能性DC。②SKOV3/DC融合细胞兼具两种亲本细胞的部分特性,但其体外增殖活性明显降低,失去亲本肿瘤的恶性生长特性。本研究为将SKOV3/DC作为肿瘤疫苗用于卵巢癌主动免疫治疗的研究提供了前期资料。Objective To prepare hybrid cells （SKOV3/DC） by fusion of dendritic cells （DC） derived from cord blood and SKOV3 ovarian cancer cells; and analyze the biological characteristics and anti-tumor immunity of SKOV3/DC. Methods ① DC cells stained with PKH26 red fluorescence were fused with SKOV3 by means of polyethyleneglycol（PEG） , and the fusion cells（SKOV3/DC） were selected with flow cytometry. ② Techniques of cell culture ,flow cytometry and light microscopy were also used in characterization of growth, morphology and immunology of SKOV3/DC in vitro. Results ① The monocytes isolated from cord blood could be induced to DC. DC expressed CDla, CD80, CD86, HLA-DR, MHC- Ⅰ molecules, but not CA125 antigen. ② SKOV3 cells expressed CA125 antigen and MHC-Ⅰ molecule but not CD1a, CD80, CD86 and HLA-DR molecules.③ The DC were fused with SKOV3 cells at a ratio of 10：1 and the fusion rate was about 8.5%. The morphological character of the SKOV3/DC was similar to its parental cells. SKOV3/DC express both CA125 antigen and high levels of CD1a, CD80, CD86, HLA-DR, MHC- Ⅰ molecules. SKOV3/DC could divide and proliferate in vitro, but the proliferative activity was significantly lower as compared with SKOV3 cells. Conclusion ① DC precursors in cord blood could be induced to differentiate and mature in media containing different cytokines. ② SKOV3/DC have some characteristics of both parental cells. The ability of dividsion and proliferation is significantly decreased as compared with SKOV3 cells, and the characteristic of malignant growth of parental SKOV3 cells has been lost. These data about SKOV3/DC vaccine found a basis for the study of the immunotherapy for ovarian cancer.

关 键 词：肿瘤疫苗 树突状细胞 卵巢癌 融合细胞 抗肿瘤免疫 

分 类 号：Q781[生物学—分子生物学]