细胞因子诱导的杀伤细胞的生物学特性研究  

The biological characteristics of cytokine-induced killer cells

作　　者：高维实[1] 闵军[1] 褚忠华[1] 陈涛[1] 魏菁[2] 区庆嘉[1] 

机构地区：[1]中山大学附属第二医院肝胆外科,广东广州510120 [2]中山大学附属第二医院医学研究中心,广东广州510120

出　　处：《中国病理生理杂志》2003年第7期946-949,T006,共5页Chinese Journal of Pathophysiology

基　　金：中山大学附属第二医院重点基金资助项目(2 0 0 2 -Z0 6 )

摘　　要：目的 :探讨细胞因子诱导的杀伤细胞 (CIK)的生物学特性。方法 :健康人非贴壁单个核细胞用含IFN-γ、IL - 1β、IL - 2、CD3单抗诱导CIK细胞。以LAK细胞作为对照。流式细胞仪和免疫细胞化学染色检测细胞表型特征 ;乳酸脱氢酶释放法分析细胞毒活性。结果 :诱导 2周后 ,CIK细胞的增殖率达到高峰 ,CD3+ 细胞占 95 %以上 ;第 3周细胞生长进入平台期。诱导 15d时 ,CD3+ CD5 6 + NKT亚群占 16 .5 % ,比例在 2 - 4周区间内无明显变化。LAK细胞增殖缓慢 ,显著低于同期CIK细胞增殖率 (P <0 .0 1)。不同效 :靶比例CIK细胞对肝癌细胞BeL - 74 0 2的特异性溶解率显著高于LAK细胞 (P <0 .0 1)。免疫细胞化学染色结果显示 ,CIK细胞高度表达HLA -DR和CD5 4抗原 ,NKT细胞体积较CD3+ CD5 6 -细胞略大 ,细胞表面有大量伪足。结论 :CIK细胞具有高度增殖能力 ,其体外杀瘤活性明显优于LAK细胞。诱导 14 - 2 1d ,CIK细胞增殖率和CD3+ CD5 6 + 阳性细胞率均达到高峰 。AIM: To investigate the biological characteristics of cytokine-induced killer (CIK) cells in vitro METHODS: The non-adhere peripheral blood monoclear cells from healthy donors were induced into CIK cells in the presence of IFN-γ, IL-1β, IL-2 and anti-CD3 antibody. LAK (lymphokine activated killer) cells were prepared as a control. The cellular phenotype were detected by FCM and immunocytochemistry and the cytotoxicity was measured by LDH release assay. RESULTS: After 2 weeks of induction, the proliferation rate of CIK cells reached a peak and the proportion of CD3 + population was above 95%, and then the cells growth entered to plateau phase at week 3. The proportion of CD3 +CD56 + NKT subset cells was 16 5% on day 15 and it had no obvious variety between 2 and 4 weeks. Correspondingly, LAK cells grew slowly and had lower proliferation rate compared with the CIK cells ( P <0 01). CIK cells showed higher specific lysis rates to BeL-7402 hepatoma cells than those of LAK cells at different effector to target ratio ( P <0 01). Immunocytochemical staining showed the CIK cells highly co-expressed HLA-DR and CD54 antigens. The NKT cells were slightly bigger than CD3 +CD56 - cells and a large quantity of pseudopodia were observed on their surface. CONCLUSION: The CIK cells have higher proliferation potency and stronger cytotoxicity to lyse tumor cells than LAK cells in vitro. Within the span of time from 14 to 21 days, the proliferation rate and the proportion of CD3 +CD56 + subset of CIK cells all reach peaks. Therefore, CIK cells in this period are suitable for clinical application.

关 键 词：细胞因子类 杀伤细胞 淋巴因子 肝肿瘤 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]