抗CD_3单克隆抗体激活杀伤细胞高效扩增培养体系的建立及细胞毒活性检测  

Establisment of anti-CD_3 monoclonal antibody activated killer cells proliferation system and cytotoxic activity

作　　者：张赢予[1] 张馨木[1] 王建秋[1] 张磊[1] 焦平[1] 王丁丁[1] 王文加[1] 马杰[1] 颜炜群[1] 

机构地区：[1]吉林大学再生医学科学研究所生物化学研究室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2007年第2期265-268,共4页Journal of Jilin University：Medicine Edition

基　　金：国家高技术研究发展计划(863计划)项目资助课题(2004AA205020)

摘　　要：目的：建立从20mL外周血中高效扩增抗CD3单克隆抗体激活的杀伤细胞（CD3AK）的培养体系,并进行体外细胞毒活性检测。方法：采用抗CD3单克隆抗体（Anti-CD3mAb）激活人外周血单个核细胞（PBMC）,在人重组白细胞介素2（rIL-2）的协同活化下,获得CD3AK。对CD3AK进行长期体外培养,进行增殖动力学、免疫表型和体外抗肿瘤活性的分析。结果：在培养第7-22天,CD3AK增殖倍数明显提高,最高可达317倍;免疫表型分析显示,CD3^＋细胞从活化前的51.9%增加至94.6%,CD4^＋细胞从19.5%增加至51.1%,CD8^＋细胞从22.1%增加至52.1%;效靶比20∶1组和40∶1组的杀伤肿瘤细胞活性明显高于10∶1组（P〈0.05和P〈0.01）;在7-19 d的培养天数之内,杀伤肿瘤细胞活性最强。结论：从外周血中高效扩增的CD3AK是以CD3^＋、CD4^＋和CD8^＋细胞为主的异质性细胞群,在体外有明显的杀伤肿瘤细胞活性。Objective To establish anti-CD3 monoclonal antibody activated killer cells （CD3AK） culture system from 20 mL peripheral blood and observe the cytotoxity against tumor cells in vitro . Methods Anti-CD3 monoclonal antibody and recombinant rIL-2 costimulated human peripheral blood mononuclear cells （PBMC） for inducing CD3AK. CD3AK was obtained and cultivated in vitro for a long time. The biological characteristics, proliferative kinetics, cytotoxicity against tumor cells and the phenotype changes of CD3AK were analyzed. Results In the cultural period of 7 - 22 d, the proliferative multiple of CD3AK increased significantly and outstandingly up to 317 times. Immunophenotypic analysis showed CD3^＋ cells increased from 51.9% before the activation to 94.6 ~. CD4^＋ cells increased from 19.5% to 51.1%. CD8^＋ cells increased from 22.1% to 52. 1%. In addition, the cytotoxicity activity of CD3AK was more significant with E/T ratio of 20 = 1 group and 40 = 1 group than 10 ： 1 group. The best antineoplasmic activity time was in the period of 7-9 d. Conclusion CD3AK, which is induced from peripheral blood, is a heterogeneity group of CD3^＋, CD4^＋ and CD8^＋ T cells and has significant antineoplasmic activity in vitro.

关 键 词：杀伤细胞 淋巴因子激活 细胞培养 抗肿瘤活性 

分 类 号：R73-36[医药卫生—肿瘤]