›› 2012, Vol. 24 ›› Issue (2): 121-124.doi: 10.3969/j.issn.1004-616x.2012.02.009

• 论著 • 上一篇    下一篇

免疫杀手细胞的体外扩增方法及其对肝癌细胞BEL-7402 的杀伤作用

王生育，罗芳洪，倪二茹，吕海燕，支黎明，颜江华，张顺浪   

  1. 1. 厦门大学医学院抗癌研究中心，福建 厦门 361005；2. 厦门长春堂科技有限公司，福建 厦门 361005
  • 收稿日期:2011-07-06 修回日期:2011-09-15 出版日期:2012-03-30 发布日期:2012-03-30
  • 通讯作者: 张顺浪

Evaluation of a modified culture method for immune-killer cells and therapeutic efficiency on hepatocarcinoma cell line BEL-7402

  • Received:2011-07-06 Revised:2011-09-15 Online:2012-03-30 Published:2012-03-30
  • Contact: ZHANG Shun-lang

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摘要： 目的： 探讨免疫杀手细胞 (immune-killer cells，IKCs)的体外扩增方法及其对人肝癌细胞BEL-7402的杀伤作用。方法：改良自然杀伤细胞培养扩增法扩增IKCs细胞，流式细胞仪分析IKCs表型，台盼蓝染色计数法观察细胞增殖状况，Alamar Blue法观察IKCs细胞对BEL-7402细胞的杀伤作用，用BALB/c裸鼠模型检测IKCs细胞对肝癌动物模型的治疗作用。结果：IKCs细胞培养14 d的细胞总数扩增300倍以上，主要由CD3+CD56+ 、CD3-CD56+和CD8+细胞亚群组成，CD3+CD56+和CD3-CD56+ 比例达85%以上，CD8+比例达到77%以上。IKCs细胞对肝癌BEL-7402细胞的杀伤实验显示，当效靶比为25∶1时和100∶1时，其杀伤活性分别为66.5%和97.8% (P＜0.01)。裸鼠肝瘤模型治疗实验显示，IKCs细胞对早期和晚期人肝癌BEL-7402细胞裸鼠模型均有显著的治疗作用，与对照组比较，其抑瘤率达69%以上 (P＜0.01)。结论：本实验建立的肿瘤免疫杀手细胞IKCs的培养扩增方法是可行的、有效的，有望成为一种新的肿瘤免疫细胞治疗方法。

关键词: 免疫杀手细胞, 肝癌, 杀伤

Abstract: OBJECTIVE: To evaluate the modified culture method and the therapeutic efficiency of immune-killer cells (IKCs) on a hepatocarcinoma cell line BEL-7402. METHODS：IKCs were induced and amplified from PBMC cells in a modified NK culture method. The immunophenotype changes were analyzed by flow cytometry. The proliferation of the cells were evaluated by Trypan-blue staining. The killing activities on BEL-7402 cell line were detected by Alamar Blue staining and their effect on BEL-7402 tumor in the nude mice model were examined by monitoring tumor growth. RESULTS：The IKCs cultured for 14 days multiplied approximately 300 times. Cell subsets of CD3+CD56+，CD3-CD56+ and CD8+ were the major components. Both ratios of CD3+CD56+ and CD3-CD56+ were higher than 85%，while the ratio of CD8+ was higher than 77%. At the effector-target ratio of 25：1 and 100：1，the killing activity against BEL-7402 cells of IKCs was 66.5% (P＜0.01) and 97.8% (P＜0.01)，respectively. Treatment with IKCs on BEL-7402 tumor in nude mice showed a noticeable higher therapeutic effect than that of control，with an inhibition ratio on tumor growth of 69% (P＜0.01). CONCLUSION：The modified culture method for IKCs was practicable and effective. The research might provide experimental basis for clinical immunotherapy of IKCs on hepatocarcinoma.

Key words: immune-killer cells, hepatocarcinoma, killing activity

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