阿糖胞苷增强白血病细胞B7分子表达及促进双功能抗体对靶细胞的杀伤  

Cytarabine enhances B7 expression on leukemic cells and promotes cytotoxic effect of bispecific antibody against target cells

作　　者：杨铭[1] 范冬梅[1] 高瀛岱[1] 赵英新[1] 周圆[1] 许元富[1] 纪庆[1] 王金宏[1] 熊冬生[1] 杨纯正[1] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室,天津300020

出　　处：《中国肿瘤生物治疗杂志》2009年第5期447-451,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.30701012);天津市科技计划资助项目(No.08ZCKFSH04100);天津市应用基础研究计划资助项目(No.07JCZDJC04900)~~

摘　　要：目的:研究阿糖胞苷(cytarabine,Ara-C)对白血病细胞共刺激分子B7表达的影响,以及联合双功能抗体anti-CD3/anti-Pgp介导T细胞对耐药白血病细胞的杀伤作用。方法:应用流式细胞术检测白血病细胞株K562和多药耐药白血病细胞株K562/A02细胞经Ara-C刺激不同时间后B7-1、B7-2分子的表达,RT-PCR方法检测B7-1mRNA、B7-2mRNA的表达,MTT法检测经Ara-C刺激的K562和K562/A02细胞对T淋巴细胞增殖的影响。CytoTox96非放射性细胞毒性分析检测Ara-C联合anti-CD3/anti-Pgp微型双功能抗体对人外周血淋巴细胞杀伤K562和K562/A02靶细胞的影响。结果:经Ara-C刺激的K562和K562/A02细胞B7-1、B7-2分子的表达较对照组明显升高;MTT结果显示,经Ara-C刺激的K562和K562/A02细胞能促进T淋巴细胞增殖;Ara-C联合anti-CD3/anti-Pgp双功能抗体在0.39:1～25:1效靶比范围内,随着效靶比的升高,介导T淋巴细胞对K562和K562/A02细胞的杀伤率随之提高,对高表达Pgp的耐药K562/A02细胞尤为明显。结论:Ara-C可上调白血病细胞B7分子的表达,从而增强anti-CD3/anti-Pgp双功能抗体介导的T细胞对靶细胞的体外杀伤作用。Objective：To observe the effects of cytarabine （Ara-C） on B7 expression on leukemia cells, and to study the effects of anti-CD3/anti-Pgp bispecific antibody on the cytotoxicity of T cells against drugresistant leukemia cells. Methods：The expressions of B7-1 and B7-2 on K562 （leukemia cells） and K562/A02 cells （drugresistant leukemia cells） were examined by flow cytometry after treatment with AraC for different periods. B7-1 and B7-2 mRNA expressions in K562 and K562/A02 cells were detected by RTPCR. The proliferation of T lymphocytes stimulated by AraCtreated K562 and K562/A02 cells was detected by MTT assay. In vitro cytotoxicity of T lymphocytes against K562 and K562/A02 cells was analyzed using CytoTox 96 nonradioactive method after treatment with anti-CD3/anti-Pgp bispecific antibody and AraC. Results：Compared with untreated cells, B7-1 and B7-2 expression on AraCtreated K562 and K562/A02 cells was significantly enhanced. MTT results showed that AraCtreated K562 and K562/A02 cells increased the proliferation of T lymhocytes. AraC combined with antiCD3/antiPgp bispecific antibody enhanced the cytotoxicity of T cells against K562 and K562/A02 target cells （T∶target, 0.39∶125∶1）, especially when against Pgp positive drugresistant K562/A02 leukemia cells. Conclusion： Ara-C can upregulate B7 expression on leukemia cells, and when combined with anti-CD3/anti-Pgp bispecific antibody it can enhance the cytotoxicity of T cells against target leukemia cells in vitro.

关 键 词：阿糖胞苷 白血病细胞 anti-CD3/anti-Pgp B7-1 B7-2 T淋巴细胞 

分 类 号：R730.3[医药卫生—肿瘤] R733.7[医药卫生—临床医学]