CIK细胞体外杀伤人肝癌细胞免疫学机制的实验研究

Study of the mechanism of cytolysis by CIK against hepatoma cells in vitro

目的 探讨CIK(cytokineinducedkiller)细胞体外杀伤人肝癌细胞的机制。方法 取健康志愿者的成分血 ,用淋巴细胞分离液密度梯度离心法分离获得外周血单个核细胞 (PBMC) ,采取本室常规方法培养诱导成CIK细胞。体外诱导CIK杀伤人肝癌细胞 ,电镜下观察肝癌细胞的形态特征改变 ;用TUNEL法检测肝癌细胞的凋亡 ;免疫细胞化学法检测并比较FasL在单个核细胞和CIK细胞表面的表达 ;用MTT法检测抗FasL单克隆抗体对CIK细胞杀伤肝癌细胞活性的影响。结果 电镜观察CIK细胞能促进细胞凋亡超微结构的改变 ,表现为细胞固缩、微绒毛脱失、核染色体边集、核固缩 ;TUNELLI的分析显示 :CIK作用后肝癌细胞凋亡指数明显增高 ;FasL在CIK细胞表达增加 ;抗Fas单抗可抑制CIK杀伤肝癌细胞的活性。结论 免疫活性细胞CIK能诱导肝癌细胞的凋亡 ,Fas FasL途径在其中起一定的作用。

Objective To investigate the mechanism of cytolysis by CIK against hepatoma cells in vitro .Methods CIK cells were induced by culturing PBMC in the completed medium containing some cytokines; ultrastructural observation and in situ DNA labeling apoptotic detection were performed; effects of anti FasmAb and expression of FasL were individually determined by MTT and immunocytochemistry (ICC) analysis.Results Electron microscopic observations showed that CIK cells could induce the hepatoma cells to apoptosis.Apoptotic label index of the hepatoma cells and FasL expression on CIK increased.Cytotoxicity was blocked after addition of anti FasmAb.Conclusion CIK cells an induce the apoptosis of hepatoma cells,and Fas/FasL pathway plays an important role in apoptosis of hepatoma cells by CIK.