负载肺癌A549抗原的树突状细胞对细胞因子诱导的杀伤细胞特异性激活研究  

Cytokine-induced Killer Cell Activation by Dentritic Cells Loaded with the Tumor Antigen from Human Lung Cancer Cell Line A549

作　　者：苏征[1] 张灏[1] 李欣[2] 汤华[2] 

机构地区：[1]天津医科大学基础医学院实验中心,天津300070 [2]天津医科大学生命科学中心,天津300070

出　　处：《中国慢性病预防与控制》2008年第1期23-26,共4页Chinese Journal of Prevention and Control of Chronic Diseases

基　　金：天津市重点攻关项目资助(023111511-8,033181988)

摘　　要：目的观察负载肺癌A549抗原的树突状细胞(DC)对细胞因子诱导的杀伤细胞(CIK)特异性肿瘤杀伤活性的作用。方法由正常人富含淋巴细胞白膜分离、细胞因子诱导制备DC细胞和CIK细胞。用流式细胞术分析DC细胞和CIK细胞的表型。经混合淋巴反应和体外细胞毒性实验观察负载A549抗原的DC对CIK具有特异性激活作用。结果表型分析显示,CIK细胞以CD3+CD56+自然杀伤细胞为主,成熟DC细胞高表达CD40、CD80、CD86和HLA-DR,并可刺激CIK细胞的增殖诱导混合淋巴反应,而负载A549抗原的DC细胞能显著增强CIK细胞特异性杀伤靶细胞的能力(P<0.05)。小鼠注射CIK、DC·CIK和Ag·DC·CIK后的肿瘤直径分别为(65.34±12.33)、(70.17±13.26)和(36.79±9.19)mm,后者的抗肿瘤作用较高(P<0.05)。结论抗原负载的DC细胞可活化CIK细胞,并使其具有特异性杀伤肿瘤细胞的能力。Objective To explore the specific anti-tumor effect of cytokine-induced killer cell （CIK） activation by dentritic cells （DC） loaded with the tumor antigen from human lung cancer cell line A549. Methods CIKs and DCs derived from the health donor＇s peripheral blood monocyte （PBMC） were isolated and induced by the cytokine in vitro. DCs were generated by culture of adherent cells from PBMC for 7days in the presence of IL-4 and GM-CSF. CIKs were generated by culture of non-adherent cells for 7 or 14 days in the presence of INF-γ IL-2, IL-1α, and mouse anti-human CD3 monoclonal antibody. The phenotype of DC and CIK was analyzed by FCM. The activity of DC was evaluated by MLR; and the cytotoxicity of CIK was assayed by MTT. The CIK expression profile of cytokine and signal transduction genes were detected by the DNA micro-array under different circumstances. Results Phenotypic analysis indicated that CD3^＋CD56^＋NKT cells were predominant in the CIK cells generated in the current study, and that DC cells expressed high levels of CD40, CD80, CD86 and HLA-DR. As demonstrated by MTr assay, the CIK cells proliferated rapidly, and the DC cells were able to induce an MLR. Both antigen-pulsed and unpulsed DC stimulated the proliferation of CIK cells, and no significant difference was found between the two kinds of DC cells. Unpulsed DC cells did not enhance cytotoxicity mediated by CIK cells even though they were able to stimulate CIK proliferation. Antigen-pulsed DC, however, stimulated CIK cells to specifically kill target cells. The specific antitumor effect was also observed in nude mice bearing tumors. Conclusion One of the major effects of antigen-pulsed DC is to activate CIK cells and to make them specific in killing tumor cells.

关 键 词：树突状细胞 细胞因子诱导的杀伤细胞 激活 

分 类 号：R73[医药卫生—肿瘤]