抗CD3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

doi:国家高技术研究发展计划(863计划）项目资助

抗CD₃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

张赢予，张馨木,王建秋，张磊，焦平，王丁丁，王文加，马杰，颜炜群

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

收稿日期:2006-07-03 修回日期:1900-01-01 出版日期:2007-03-28 发布日期:2007-03-28
通讯作者: 颜炜群

Establisment of anti-CD₃ monoclonal antibody activated killer cells proliferation system and cytotoxic activity

ZHANG Ying-yu, ZHANG Xin-mu,WANG Jian-qiu,ZHANG Lei,JIAO Ping,WANG Ding-ding,WANG Wen-jia,MA-Jie,YAN Wei-qun

Department of Biochemistry,Institute of Frontier Medical Sciences,Jilin University，Changchun 130021,China

Received:2006-07-03 Revised:1900-01-01 Online:2007-03-28 Published:2007-03-28
Contact: YAN Wei-qun

摘要/Abstract

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(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-CD₃ mAb）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。

关键词: 淋巴因子激活, 细胞培养, 抗肿瘤活性

Abstract: 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.

Key words: lymphokine-activated, cell culture, cytotoxicity against tumor cells

中图分类号:

R73-36

张赢予，张馨木,王建秋，张磊，焦平，王丁丁，王文加，马杰，颜炜群. 抗CD₃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[J]. J4, 2007, 33(2): 265-268.

ZHANG Ying-yu, ZHANG Xin-mu,WANG Jian-qiu,ZHANG Lei,JIAO Ping,WANG Ding-ding,WANG Wen-jia,MA-Jie,YAN Wei-qun. Establisment of anti-CD₃ monoclonal antibody activated killer cells proliferation system and cytotoxic activity[J]. J4, 2007, 33(2): 265-268.

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抗CD₃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-CD₃ monoclonal antibody activated killer cells proliferation system and cytotoxic activity

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