大肠癌肿瘤抗原修饰的树突状细胞活性检测

目的:测定经大肠癌肿瘤抗原修饰的树突状细胞(DC)的活性. 方法:分离结肠癌患者的外周血单个核细胞,应用粒细胞/ 巨噬细胞集落刺激因子联合白细胞介素-4进行体外诱导,获取DC,以流式细胞仪进行表型鉴定;用Lovo细胞制备大肠癌肿瘤抗原并用以修饰DC,以肿瘤抗原修饰后的DC为刺激细胞,以自体淋巴细胞为效应细胞,采用MTT法检测刺激细胞与效应细胞在不同比例(1:10,1:100,1:1 000)以及不同共孵时间(24 h,48 h,72 h)下,效应细胞对大肠癌细胞的排斥杀伤率.结果:从大肠癌患者外周血单个核细胞中成功诱导出表达CD11c、CD80和HLA-DR的DC,经肿瘤抗原修饰后,上述分子表达上调.肿瘤抗原修饰后的DC与淋巴细胞以1:100共孵48 h时,淋巴细胞的肿瘤细胞排斥杀伤率较高.结论:从大肠癌患者外周血中分离诱导出的DC,被肿瘤抗原修饰后,在体外可激活自体淋巴细胞,提高淋巴细胞的杀瘤活性.

Activity of dendritic cells modified by tumor antigen

Aim: To study the activity of dendritic cells (DC) induced by peripheral blood mononuclear cells (PBMCs) of colony carcinoma patients and modified by tumor antigen.Methods:Human PBMCs from the colony carcinoma patient were induced using recombinant human granulocyte macrophage colony stimulating factor, interleukin-4 combinedly and were stimulated with whole cell lysate of Lovo cell lines to generate DC modified by tumor antigens,and then the phenotype of the DC was identified by flow cytometer. The cytotoxicity of lymphocytes by DC modified by tumor antigens to Lovo cells were assayed with MTT method. Results: DC derived from human of colony carcinoma patients expressed CD11c, CD80, HLA-DR, and the expressions upgraded as DC modified by tumor antigens. The activity of lymphocytes excited by DC modified by tumor antigen at the ratio that DC and lymphocytes was 1:100 and that hatch time was 48 h, the rate of the repelling and killing of the tumor cells of the lymphocytes were higher. Conclusion: DC, derived from human PBMCs and modified by tumor antigens, could excite the autologous lymphocytes.