中华眼镜蛇膜毒素与McAb偶联物对白血病细胞的杀伤作用  

Specific Killing of Human Leukemic T-Cells by a Monoclonal Antibody Coupled to the Cytotoxin from Chinese Cobra

作　　者：褚嘉佑[1] 张和君[1] 杨爱德[2] 王辨明 何其伟[3] 胡忠[4] 朱晓梅[4] 龚春梅[1] 郭仁[1] 

机构地区：[1]中国医学科学院医学生物学研究所 [2]同济医科大学血液病研究所 [3]中国科学院昆明动物研究所 [4]中国科学院昆明植物研究所

出　　处：《中国医学科学院学报》1992年第3期179-184,共6页Acta Academiae Medicinae Sinicae

摘　　要：用中华眼镜蛇膜毒素(CT)与抗人T细胞McAb Wu71偶联制备免疫毒素Wu71:CT。经SDS-聚丙烯酰胺凝胶电泳、免疫双扩散和间接免疫荧光检测,证实结合物中McAb和CT均保持活性。在无Ca^(2+)、Mg^(2+)的培养基中将人白血病T淋巴细胞系CEM细胞与Wu71:CT共同孵育,先加入Ca^(2+)以抑制CT的溶细胞作用,而不影响McAb与靶细胞的结合;一定时间后用Mg^(2+)和金属螯合剂EGTA恢复CT的溶胞作用,结果在Wu71:CT 0.5×10^(-6)mol/L浓度下靶细胞CEM 82.4％被杀伤,而抗原阴性的非靶细胞Raji仅死亡23.3％,表明Wu71:CT有较好的特异性杀伤作用。扫描电镜显示,免疫毒素作用下的CEM细胞膜破裂、萎缩、崩溃,而非靶细胞大部分保持完整。提示McAb与CT偶合物在白血病和肿瘤的导向治疗中可能具有潜在价值。The immunotoxin (IT) was prepared by conjugating cytotoxin from Chinese Cobra (Naja naja Atra) venom with monoclonal antibody (McAb) Wu7l directed to human T-cells. First, calcium ions were used to suppress the cytolytic reaction while cell antigen was allowed to react with McAb. Then magnesium ions and the chelating agent EGTA were used to abolish the calcium inhibition. With this treatment, the IT showed high cytotoxicity for the leukemic cell line CEM, which was antigen positive (82.4% of the cells were killed at a concentration of 0.5× 10-8mol/L of IT), but very little cytotoxicity for the antigen negative cell line Raji (23.3% cells were killed at the same concentration). Under scanninng electron microscopy, it could be seen that the cell membranes of CEM were broken by IT, and the cells had died. The results suggest that the McAb Wu71 plus cytotoxin IT has potential application in leukemia therapy.

关 键 词：免疫毒素 蛇膜毒素 白血病 

分 类 号：R733.705[医药卫生—肿瘤]