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目的:观察抗死亡受体5(DR5)单克隆抗体(mAb)-mDRA-6对白血病细胞的凋亡作用。方法:DR5蛋白免疫BALB/c小鼠,融合筛选抗DR5杂交瘤细胞,制备抗人DR5mAb-mDRA-6;荧光显微镜下观察mDRA-6作用下白血病细胞Jurkat、HL-60、K562的形态变化;MTT法测定不同浓度的mDRA-6对Jurkat、HL-60、K562细胞存活率的影响;通过FITC-Annexin V及PI标记细胞,以流式细胞术检测mDRA-6对Jurkat、HL-60、K562细胞凋亡率的影响。结果:mDRA-6导致Jurkat、HL-60细胞染色质浓缩、断裂,细胞出芽,凋亡小体形成;mDRA-6对Jurkat、HL-60细胞具有明显的杀伤作用,但对K562的杀伤作用较弱,25mg/L的mDRA-6作用12h,Jurkat、HL-60、K562细胞死亡率分别为88.76%,59.76%,5.18%。Annexin V及PI双染显示1mg/L的mDRA-6作用10h,Jurkat细胞凋亡率为86.06%,10mg/L的mDRA-6作用10h,HL-60细胞凋亡率为48.11%,但10mg/L的mDRA-6作用K562细胞10h,细胞凋亡不明显。结论:抗DR5mAb mDRA-6能够诱导白血病细胞凋亡,不同的白血病细胞株对mDRA-6的敏感性不同。
AIM: To observe the apoptosis of leukemia cells by anti-death receptor 5 (mAb) -mDRA-6 monoclonal antibody (mAb). Methods: BALB / c mice were immunized with DR5 protein and anti-DR5 mAb-mDRA-6 was prepared by fusion screening of anti-DR5 hybridoma cells. Morphological changes of Jurkat, HL-60 and K562 leukemia cells were observed under fluorescence microscope. The effect of different concentrations of mDRA-6 on the survival rate of Jurkat, HL-60 and K562 cells was determined by MTT assay. The effect of mDRA-6 on Jurkat, HL-60 and K562 cells was detected by FITC-Annexin V and PI staining Effect of apoptosis rate. Results: mDRA-6 resulted in chromatin condensation, rupture, sprouting and apoptotic body formation in Jurkat and HL-60 cells. MDRA-6 had a significant killing effect on Jurkat and HL-60 cells, but the killing effect on K562 was stronger than mDRA-6 The cell death rates of Jurkat, HL-60 and K562 cells were 88.76%, 59.76% and 5.18% respectively after being treated with 25mg / L mDRA-6 for 12h. Annexin V and PI double staining showed that the apoptotic rate of HL-60 cells was 48.11% when 10 mg / L mDRA-6 was treated with 1 mg / L mDRA-6 for 10h, the apoptosis rate of Jurkat cells was 86.06% and 10 mg / L mDRA-6 effect K562 cells 10h, apoptosis was not obvious. CONCLUSION: Anti-DR5 mAb mDRA-6 can induce apoptosis of leukemia cells. Different leukemia cell lines have different sensitivity to mDRA-6.