摘要
目的:探讨CD137信号对CIK细胞的增殖和功能调节作用。方法:分离健康志愿者外周血单个核细胞(PBMCs),实验组在常规CIK培养体系中加入CD137单抗(CD137-CIK组),对照组在常规CIK培养体系中加入鼠IgG1同型对照(IgG1-CIK组)。苔盼蓝拒染法细胞计数分析细胞增殖;流式细胞术检测细胞表型及细胞内因子的表达;LDH酶释放法检测CIK细胞杀伤活性。结果:CD137-CIK组细胞体外扩增效率显著高于IgG1-CIK组,CD137-CIK组细胞浓度最高达(9.87±0.57)×106/ml;IgG1-CIK组最高为(7.02±0.68)×106/ml;CD137-CIK组和IgG1-CIK组细胞中CD3+CD56+细胞比例至28天分别达到(39.86±4.69)%和(29.14±5.12)%(P<0.05);经CD137mAb作用后,其体外杀伤肺癌细胞株A549活性明显高于对照组(P<0.05);第0、7、14、21天流式检测IFN-γ表达,实验组CD3+CD56+细胞IFN-γ的表达显著高于对照组。结论:CD137mAb介导的共刺激信号可以促进CIK细胞的体外增殖活性,并增强CIK细胞体外抗瘤作用。其中CD137-CIK组CD3+CD56+细胞的比例及其IFN-γ表达显著提高,这可能是CD137信号增强CIK抗瘤作用的重要原因。
Objective:To investigate the effect of CD137 signal on regulation of CIK cell proliferation and function. Methods:CIK cells were induced from peripheral blood mononuclear cells in the culture with the presence of CD3mAb, IL-2,IFN-γ Cells treated with CD137mAb or mouse IgG1 isotype control was assigned to experimental group or control group, separately. The cell proliferation was determined by cell counting with trypan blue staining test. Phenotypes and IFN-γ production of CIK cells were analyzed by flow cytometry (FCM) ;Cytotoxicity was measured by LDH activity released from the supernatant of cell cultures. Results:The concentration of CD137-CIK cells in experimental group was dramatically higher than traditional CIK cells in control group. The concentration was respectively arrived at (9.87 ± 0.57)x10^6/ml and (7.02 ± 0.68)xl0^6/ml;The percentages of CD3^+CD56^+ cells and the expression of IFN-γ in experimental group were increased significantly compared with control group as well;The CD137-CIK cells possessed much higher ability to kill lung cancer cell(A549) than IgG1-CIK cells did. Conclusion:CD137-CIK cells exhibited strong activities of proliferation and cytotoxicity against A549 cells.
出处
《南京医科大学学报(自然科学版)》
CAS
CSCD
北大核心
2008年第2期149-153,239,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助项目(30772549)