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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 study the in vitro expansion and culture of dendritic cells (DCs) derived from cord blood and to induce specific anti-ovarian cancer cell immune responses. Methods ① After mononuclear cells were isolated from umbilical cord blood, monocytes (Mo) were obtained. Differentiation and expansion were induced by granulocyte-macrophage colony-forming agent (GM "CSF), IL-4, colony stimulating factor (SCF) and tyrosine kinase receptor III ligand 7d after the phenotypic analysis by flow cytometry; ② induction of monocyte differentiation on the 3rd day adding human ovarian cancer cell line SKOV3 frozen-thawed antigens, co-cultured for 4 days to obtain tumor antigen-loaded mature DC; will be sensitized DC and allogeneic T lymphocytes isolated from cord blood were co-cultured for 6 days to obtain effector cells; Umbilical cord blood-derived Mo was differentiated into mature DCs after 7 days by cytokines and highly expressed specific antigens such as CD83, CD86, CD80, and CD1a; ② DCs could be loaded and presented with tumor antigens to induce tumor-specific cytotoxic T lymphocytes (CTL). Different concentrations of CTL had specific cytotoxic and inhibitory effects on ovarian cancer cell line SKOV3 (P <0.05). CONCLUSION: DC precursors (ie, mononuclear cells) in cord blood can differentiate and proliferate in vitro to mature functional DCs and induce specific immune responses against ovarian cancer cells, indicating a promising clinical application , Convenient and effective method of tumor immunotherapy.