抗原致敏树突状细胞诱导CIK对肺癌细胞杀伤作用的研究

Effect of Cytokine-induced Killer Cells Induced by Dendritic Cells Pulsed with Tumor Antigen on Cytotoxicity Against Lung Cancer Cell

[目的]研究肿瘤抗原致敏的树突状细胞(DC)诱导淋巴因子激活的杀伤细胞(LAK)和细胞因子诱导的杀伤细胞(CIK)对肺癌细胞株A549和肺腺癌原代细胞的杀伤作用。[方法]取健康人外周血单个核细胞,常规诱导出DC、CIK、LAK细胞。用肺癌A549细胞提取的肿瘤抗原冲击DC,倒置显微镜下观察DC形态。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。LDH释放法测定杀伤活性。[结果]DC经肿瘤抗原冲击后在镜下呈典型成熟形态,其表面分子CD40、CD80、CD86和HLA-DR的表达明显较未经肿瘤抗原冲击的DC高。DC+CIK细胞对A549和肺腺癌原代细胞的杀伤活性高于CIK细胞、LAK细胞和DC+LAK细胞(P<0.05),随着效靶比的升高,其杀伤效应随之增强(P<0.05)。[结论]肿瘤抗原致敏的DC可诱导特异性CIK细胞,DC+CIK细胞对A549和肺腺癌原代细胞的杀伤作用明显高于DC+LAK、CIK、LAK细胞。

[Purpose]To investigate the lethal effect of dendritic cells （DCs） allergized by tumor antigen combining with cytokine-induced killer （CIK） cells or lymphokine-activated killer （LAK） cells on A549 cells and primary cells of adenocarcinoma of lung. [Methods] PBMCs were isolated from healthy donors and DC, CIK and LAK cells were induced by routine methods. DCs were pulsed with tumor antigen extracted from A549 cell, and the morphs of DCs were observed by inverted microscopy. Phenotypes of DCs were analyzed by flow cytometry after being pulsed with tumor antigen or not. The cytotoxic activity was detected by LDH release assay. [Results] DCs became obviously mature after being pulsed with tumor antigen, and their expressions of surface molecules （CD40, CD80, CD86 and HLA-DR） were much higher than those of DCs not being pulsed. The cytotoxic effect of DC＋CIK cells on A549 cells and primary cell of lung adenocarcinoma was significantly higher than that of CIK cells, DC＋LAK cells and LAK cells（P〈0.05）, and the cytotoxic effect was enhanced with the increasing ratio of effector cells to target cells（P〈0.05）.[Conclusion] DCs allergized by tumor antigen can induce specific CIK cells which can significantly enhance cytotoxic activity on A549 cell strain and primary cells of lung adenocarcinoma. DC＋CIK cells show stronger cytotoxic effect than DC＋LAK, CIK and LAK cells.