阿糖胞苷对抗CD3／抗P—gp双功能抗体介导的T淋巴细胞免疫杀伤多药耐药白血病细胞的影响  

The influence of Ara-C on anti-CD3/ anti-Pgp mediating T-lymphocytes activities against multi-drug resistant leukemia cells

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

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：《中华血液学杂志》2009年第12期812-815,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金（30701012）;天津市科技计划（08ZCKFSH04100）;天津市应用基础研究计划（07JCZDJC04900）

摘　　要：目的探讨阿糖胞苷对抗CD3／抗P—gp双功能抗体介导的T淋巴细胞免疫杀伤多药耐药白血病细胞的影响。方法采用抗E-tag亲和层析柱分离纯化抗CD3／抗P—gp微型双功能抗体，用阿糖胞苷刺激K562和K562／A02细胞，并用流式细胞术检测K562和K562／A02细胞B7—1、B7—2分子的表达，用CytoTox 96非放射性细胞毒试剂盒检测阿糖胞苷对抗CD3／抗P—gp双功能抗体介导的T淋巴细胞免疫杀伤K562／A02细胞的影响。结果经阿糖胞苷刺激的K562和K562／A02细胞B7-1、B7-2分子的表达较未刺激的对照组明显升高。阿糖胞苷对抗CD3／抗P—gp双功能抗体介导的T淋巴细胞在效靶比0．39：1～25：1范围内，激活T细胞对耐药白血病细胞的杀伤率为（16．44±1．20）％～（60．49±2．90）％，其杀伤率与效靶比和抗体浓度呈依赖关系（P〈0．05）。结论阿糖胞苷可明显提高抗CD3／抗P—gp双功能抗体介导的人T淋巴细胞对高表达P—gp抗原的耐药白血病细胞的杀伤效府。Objective To investigate the role of Ara-C in regulating anti-CD3/anti-Pgp mediating T- lymphocytes activities against muhi-drug resistant leukemia cells. Methods The diabody of anti-CD3/anti- Pgp was purified by E-tag affinity chromatography. K562 and K562/A02 cells were treated with Ara-C. The expressions of B7-1 and B7-2 on K562 and K562/AO2 cells were detected by FACS. The cytotoxicity of Tlymphocytes combined with anti-CD3/anU-Pgp plus Ara-C was analyzed by CytoTox 96 nonradioactive method. Results The expressions of B7-1 and B7-2 on K562 and K562/A02 cells treated by Ara-C was significantly higher than those untreated. The effect/target ratio was from 0.39：1 to 25： 1, and the killing rate of activated T cells to anti-drug-resistant leukemia cells was from（ 16.44 ± 1.20） % to （60.49 ± 2.90） %. The killing rates were increased gradually, with both the effect/target ratio and the antibody concentration increasing（ P 〈 0.05 ）. Conclusion Ara-C may be an important adjuvant for improving anti-CD3/anti-Pgp mediating T-lymphocytes activities against multi-drug resistant leukemia cells.

关 键 词：阿糖胞苷 双功能抗体 抗CD3/抗Pgp T淋巴细胞 B7分子 

分 类 号：R733.7[医药卫生—肿瘤]