摘要
目的探索一种在体外从人外周血单个核细胞(PBMC)扩增CD5+6淋巴细胞的方法。方法以干细胞培养基(SCGM)为基础培养基,设计IL-2/抗IL-2抗体(IL-2/抗IL-2抗体质量比为1/20)实验组以及IL-2+IL-15、相应因子浓度的IL-2和抗IL-2抗体三个对照组,从PBMC中诱导扩增CD5+6淋巴细胞,流式细胞仪检测第7、10天CD3/CD56表达率,四甲基偶氮唑蓝法检测CD56+细胞对肿瘤细胞株K562的杀伤活性。结果培养第7、10天时,CD56+细胞分别扩增了(24.67±3.14)和(31.63±5.01)倍,其中CD3+CD5+6和CD3-CD5+6细胞分别扩增(110.93±19.10)、(7.8±1.17)和(157.60±46.31)、(12.03±1.64)倍,实验组与对照组比较扩增倍数差别有统计学意义(P<0.05)。培养第10天,效/靶比10∶1时,实验组CD56+细胞对K562细胞的杀伤率为(83.46±1.56)%,与对照组比较具有更强细胞毒性作用(P<0.05)。结论在SCGM为基础培养基条件下,IL-2/抗IL-2抗体复合物能更有效地扩增CD5+6细胞毒性免疫效应细胞,为肿瘤过继免疫治疗提供了一种扩增外周血CD5+6淋巴细胞的新方法。
Objective To explore an efficient method for indue.：ng and expanding CD56^＋ lymphocytes from human peripheral blood mononuelear cells （PBMC） in vitro. Methods PBMCs were euhurod in stem cell growth medium （SCGM） supplemented with IL-2/anti-IL-2 complex and IL-2 ＋ IL-15 respectively to induce and expand CD56^＋ lymphocytes, the immune phenotypes were analyzed by flow cytometry （FCM） on the 7th and 10th day after incubation seperately, and the cytotoxieity to K562 targets was detected by MTT assay after culturing for 10 days. Results The CD56^＋ cells cultured with IL- 2/anti-IL-2 complex got the best expansion and expanded to 24.67 ± 3.14 folds in 7 days and 31.63 ± 5.01 folds in 10 days after incubation. Among them, the expanded folds for CD3^＋ CD56^＋ and CD3^- CD56^＋ were （ 110.93 ± 19. 10）, （7.8± 1.17 ） after culturing for 7 days and（ 157.60 ± 46.31 ）, （ 12.03±1.64） after 10 days, respectively. At 10：1 of effecter/target ratio, the cytotoxicity to K562 targets increased up to（83.46 ± 1.56）%, significantly higher than that of the other control groups. Conclusions CD56^＋ lymphocytes could be expanded effectively from PBMC using SCGM under the stimulation and induction of IL-2/anti-IL-2 complex. This work provided a novel and effective method of expanding human CD56^＋ lymphocytes in vitro for the adoptive immunotherapy of malignant tumors.
出处
《山东医药》
CAS
北大核心
2009年第13期1-3,共3页
Shandong Medical Journal
基金
青岛大学医学院附属医院"222重点项目"(2004Y2D-4)
