抗原致敏DC联合LAK细胞对肺腺癌细胞杀伤作用的研究

Research on the Effect of Dendritic Cells Combined with Lymphokine Activited Killer Cells on Cytotoxicity Against Lung Cancer Cells

目的:观察抗原致敏树突状细胞(DC)联合淋巴因子激活的杀伤(LAK)细胞对肺腺癌细胞的杀伤作用。方法:采用密度梯度离心法分离出外周血单个核细胞(PBMC),常规诱导出DC、LAK细胞。倒置显微镜观察细胞的形态和增殖情况;应用流式细胞仪(FCM)测定细胞表型上变化鉴定这两种细胞。流式细胞仪检测DC经抗原冲击和未经抗原冲击后其表型变化;利用四甲基偶氮唑蓝(MTT)检测法把DC-LAK细胞和DC-LAK-A549细胞作为效应细胞,肺腺癌A549细胞作为靶细胞,进行杀伤试验。结果:①体外诱导单个核细胞培养出DC和LAK细胞。②共培养后DC-LAK细胞的免疫表型表达增加。抗原致敏DC联合LAK细胞(DC-A549-LAK)组的免疫表型表达与对照组相比更有显著性意义。③共培养细胞在增殖倍率上有显著意义。④MTT显示共培养后的LAK细胞杀伤肿瘤细胞活性增强,抗原致敏后DC-A549-LAK抗肿瘤效应增强更显著。结论:从外周血单个核细胞中诱导出具有典型形态和免疫表型的DC和LAK细胞。DC和LAK细胞共培养细胞,增殖活性、免疫表型、杀伤活性方面高于单纯培养的LAK细胞。抗原致敏后DC-A549-LAK抗肿瘤效应增强更显著。

Objective： To observe the effect of Dendritic Cells（DC） combined with Lymphokine Aetivited Kliller（LAK） Cells on Cytotoxicity against Lung Cancer Cells. Method. Density gradient centrifugation was used to separated peripheral blood mononuclear cells（PBMC）, DC and LAK were conventional induced. Cell morphological and proliferation were observed by, inverted microscope . By FCM（flow cytometry measurement） to measure the change of phenotype and identify two kinds of cells. To detect the change of phenotype of DC by lysate antigen or no lysate antigen. Killing test was done by MTT. Result： （1）Invitro - induction PBMC cul- tured DC and LAK. （2）After co - culture, immune phenotype expression of DC - LAK was increased; Comparing to control group , there was signification different in immune phenotype expression of DC combined with LAK. （3）Co - cultured cells has significance in proliferic times. （4）MTT shows co - cultured LAK killed tumor cells ac- tivity increased, After lysate antigen, anti - tumor effect of DC - A549 - LAK markedly enhanced. Concision： With typical morphology and phenotype of DC and LAK were induced from PBMC. Co - cultured cells from DC and LAK were higher than simple culture LAK in proliferic activity, immune phenotype, killed tumor cells ac- tivity. After lysate antigen , anti - tumor effect of DC - A549 - LAK markedly enhanced.