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体外诱导抗卵巢癌肿瘤免疫的能力。方法:1)将用pKH26红色荧光染料标记脐血来源的DC后,聚乙二醇法(PEG)融合DC与人卵巢癌细胞株SKOV3细胞,流式细胞仪分选融合细胞。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;四甲基偶氮唑蓝(MTT)法观察融合细胞体外刺激混合淋巴细胞反应的能力及诱导特异性肿瘤免疫的能力。结果:1)DC和SKOV3按10:1比例融合,融合率约为8.5%。融合细胞可在体外缓慢增殖,CA125抗原及CD1a、CD80、CD86、HLA-DR、MHC-I分子表达阳性。2) SKOV3/DC在体外能有效的激发混合淋巴细胞增殖反应,可明显激活细胞毒性T淋巴细胞(CTL),对卵巢癌细胞株SKOV3有特异性杀伤效应。结论:利用PEG法制备的SKOV3/DC融合细胞兼具两种亲本细胞的部分特性,在体外能够诱导特异性抗肿瘤免疫。本研究将SKOV3/DC作为肿瘤疫苗,为其在卵巢癌主动免疫治疗研究中的进一步应用打下了基础。
Objective： To prepare hybrid SKOV3/DC cells by fusing dendritic ceils （DC） derived from cord blood and SKOV3 ovarian cancer ceils and to analyze anti-tumor immune response to ovarian cancer cells in vitro. Methods： 1 ） Using polyethylene glycol （PEG） SKOV3 ceils were fused to DC that was then stained with red fluorescent PKH26. The SKOV3/DC fused cells were selected by flow cytometry. 2） Flow cytometry and light microscopy were used to study the morphology of SKOV3/DC in vitro. MTF assay was used to detect the. fused cells＇ ability to activate a mixed lymphocyte reaction. The killing action of the specific anti-tumor immune response was also assessed. Results： 1 ） The DC were fused with SKOV3 cells at a ratio of 10：1 and the fusion rate was about 8.5%. SKOV3/DC could proliferate slowly in vitro and expressed CA125, CD1a, CD80, CD86, HLA-DR, and MHC-I molecules. 2） SKOV3/DC could stimulate a mixed lymphocyte reaction （MLR） efficiently in vitro： SKOV3/DC could activate cytotoxie T lymphocytes that could kill the SKOV3 cells specifically. Conclusion： SKOV3/DC obtained by PEG treatment have some characteristics of both parental cells and were capable of inducing a specific anti-tumor immune response to ovarian cancer cells in vitro. These data simulating an SKOV3/DC vaccine provide evidence for effective immunotherapy against ovarian cancer.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第14期789-793,共5页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:39970770)
关键词
肿瘤疫苗
树突状细胞
卵巢癌
融合细胞
抗肿瘤免疫
Tumor vaccine Dendritic cells Ovarian cancer Fused cells Anti-tumor immunity
