肿瘤可溶性抗原联合超抗原致敏的DC诱导CIK对鼻咽癌细胞杀伤作用的研究

Effect of cytokine-induced killer cells induced by dendritic cells pulsed with tumor soluble antigen and superantigen on cytotoxicity against nasopharyngeal carcinoma cell

目的探讨肿瘤可溶性抗原联合超抗原致敏的树突状细胞(dendritic cells,DCs)诱导细胞因子激活的杀伤细胞(cytokine-induced kill cells,CIK)对鼻咽癌细胞CNE-1的体外杀伤作用。方法分离人外周血单个核细胞(peripheral blood mononuclear cell,PBMC),采用GM-CSF、IL-4和TNF-α体外诱导培养DC,采用CD3Ab、IFN-γ、IL-2、IL-1等细胞因子诱导培养CIK细胞。鼻咽癌肿瘤可溶性抗原(tumor soluble antigen,TSA)联合超抗原金黄色葡萄球菌肠毒素C(staphylococcal enterotoxin C,SEC)致敏DC;将未负载抗原的DC、单纯负载鼻咽癌抗原TSA的DC、单纯负载超抗原SEC的DC及负载TSA和SEC的DC分别与CIK细胞共同孵育,诱导产生特异性的细胞毒性T细胞(cyctotoxic T lymphocyte,CTL)(各组效应细胞分别简称为DC-CIK、TSA-DC-CIK、SEC-DC-CIK、TSA-SEC-DCCIK);采用CCK-8法检测各效应细胞对靶细胞CNE-1的杀伤作用。结果成功培养出高表达HLA-DR、CD1a、CD80的DC;效靶比为50∶1时,TSA-SEC-DC-CIK对靶细胞CNE-1的杀伤率为85.46±2.12%,明显高于DC-CIK(51.36±0.61%)、TSA-DC-CIK(63.15±2.25%)及SEC-DC-CIK(65.32±3.24%)。结论肿瘤可溶性抗原联合超抗原致敏的DC诱导CIK对鼻咽癌细胞有高效特异杀伤作用。

Objective To investigate the killing effect of dendritic cells（DCs）allergized by tumor soluble antigen and superantigen combining with cytokine-induced killer（CIK）cells on human nasopharyngeal carcinoma cell lines CNE-1 in vitro.Methods PBMCs were isolated from healthy donors and DCs were induced and cultured by GM-CSF,IL-4and TNF-α,CIK cells were induced by CD3 Ab,IFN-γ,IL-2,IL-1.DCs modified by nasopharyngeal carcinoma tumor soluble antigen（TSA）and staphylococcal enterotoxin C（SEC）;DCs modified by nothing or DCs modified by TSA or DCs modified by SEC or DCs modified by TSA and SEC were cultivated together with CIK cells,and the obtained cells were named DC-CIK or TSA-DC-CIK or SEC-DC-CIK or TSA-SEC-DC-CIK as effector cells.The antitumor activity of every effector cells against target cells was assayed with CCK-8 method.Results Induced DCs expressed more CD1 a,CD80and HLA-DR.When the E：T is 50∶1,the killing ratio of the TSA-SEC-DC-CIK in vitro to CNE-1is 85.46±2.12%,which is higher than DC-CIK（51.36±0.61%）,TSA-DC-CIK（63.15±2.25%）and SECDC-CIK（65.32±3.24%）.Conclusion DCs allergized by tumor soluble antigen and superantigen can induce specific CIK cells which can significantly enhance antitumor effect on nasopharyngealcarcinoma cell.