脐血单个核细胞体外诱导树突状细胞联合CTLs抗肿瘤效应的研究

Inducement of Dendritic Cells Derived from Cord Blood Mononuclear Cells and CTLs Antitumor Immunity in Vitro

目的探讨从人脐血中定向诱导扩增树突状细胞(dendritic cells,DCs)的方法及其在抗肿瘤免疫治疗中的作用.方法常规无菌采集新鲜脐血,密度梯度离心分离脐血单个核细胞(cord b lood mononuc learcells,CBMCs),加入细胞因子(GM-CSF+IL-4+TNF-α)定向诱导培养并进行相应的表型鉴定.观察肿瘤抗原负载后诱导特异性细胞毒T淋巴细胞生成和对肿瘤细胞杀伤作用.结果从人脐血分离到的CBMCs经GM-CSF+IL-4+TNF-α共同培养后,第7天镜下即可见典型形态的DCs,细胞表型检测见CD1 a+细胞比例增加到(20.8±1.62)%.DCs负载肿瘤抗原后能诱导产生肿瘤特异性淋巴细胞,并可特异性抑制YTMLC肿瘤细胞.结论经细胞因子组合可以从脐血中诱导出DCs,并可诱导肿瘤特异性细胞毒性T淋巴细胞(cyto-toxic T-lymphocytes,CTLs)的产生,为进一步开展DCs的实验研究及临床应用奠定了基础.

Objective To investigate the way of inducing dendritic cells from precursor in human cord blood and its role in antitumor immunity. Methods Cord blood was collected under sterile condition and the cord blood mononuclear cells were separated by centrifugal in density gradient. CBMCs were cultured with GM - CSF ＋IL-4 ＋TNF- α and cell phenotype was analyzed with CD1a, CD83 antibody by using indirect immunofluorescence assays. The effects of DCs pulsed with tumoral antigens on cytotoxic T lymphocytes （CTLs） inducement and growth inhibition of YTMLC cells were assayed. Results Our results indicated that DCs precursors in human cord blood can be induced to differentiate in the medium containing GM - CSF, IL - 4 and TNF - α. The cells with typical morphological properties of DCs were observed at the 7th day. At that time, （20. 8± 1.62 ）% CD1a^ ＋ cells were obtained. After incubation with tumor cytolysis antigen, the DCs can activate the CTLs to be- come tumor specialized CTLs, which had shown significantly inhibition on growth of YTMLC tumor cell line. Conclusion The precursors in human cord blood can be induced to functional DCs which activate T lymphocyte to become tumor specialized CTLs.