抗CD47抗体联合阿糖胞苷靶向治疗NOD/SCID小鼠单核细胞白血病的研究

王颖超; 冯磊; 殷楚云; 马丽娜; 魏永纬; 王春美; 盛光耀

doi:10.7499/j.issn.1008-8830.2013.07.017

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中国当代儿科杂志 ›› 2013, Vol. 15 ›› Issue (7) : 577-582. DOI: 10.7499/j.issn.1008-8830.2013.07.017

论著·实验研究

抗CD47抗体联合阿糖胞苷靶向治疗NOD/SCID小鼠单核细胞白血病的研究

王颖超，冯磊，殷楚云，马丽娜，魏永纬，王春美，盛光耀

作者信息 +

Effectiveness of Anti-CD47 antibody and Ara-C combination targeting therapy NOD/SCID mouse of myeloid leukemia

WANG Ying-Chao, FENG Lei, YIN Chu-Yun, MA Li-Na, WEI Yong-Wei, WANG Chun-Mei, SHENG Guang-Yao

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摘要

目的：探讨CD47在急性髓细胞白血病NOD/SCID小鼠模型中的预后意义及靶向治疗的最佳策略。方法：将分选的CD34+CD38-白血病干细胞（leukemia stem cells, LSCs）移植入NOD/SCID小鼠体内，建立小鼠急性单核细胞白血病模型；抗人CD47单克隆抗体单独或联合阿糖胞苷治疗白血病小鼠7~14 d，并进行疗效分析。将LSCs与小鼠巨噬细胞在含抗CD47单克隆抗体的培养液中共培养，观察CD47对巨噬细胞吞噬LSCs能力的影响。结果：THP-1细胞中存在CD34+CD38- LSCs，比例约为0.12%±0.06%，将分选后的CD34+CD38- LSCs（比例高达97.0%±1.7%）移植入NOD/SCID小鼠后成功建立白血病模型。体内实验表明，阿糖胞苷（7 d）联合抗CD47单克隆抗体（14 d）治疗后，白血病小鼠外周血和骨髓中CD33+CD45+白血病细胞下降最明显（P<0.01），生存时间明显长于其它各组。体外共培养2 h后，抗CD47单克隆抗体组吞噬指数（76.9%±12.2%）明显高于抗CD45单克隆抗体组（7.60%±2.4%，P<0.01）。结论：CD47高表达是急性髓细胞白血病的预后不良因素。阿糖胞苷联合抗CD47单克隆抗体可有效杀灭普通白血病细胞和白血病干细胞，对彻底治愈急性髓细胞白血病具有重要临床意义。

Abstract

OBJECTIVE: To study the prognostic significance of CD47 in a NOD/SCID mouse model of acute myeloid leukemia (AML) and the best strategy for targeted therapy for this disease. METHODS: CD34+CD38- leukemia stem cells (LSCs) were separated and transplanted into NOD/SCID mice to establish a mouse model of acute monocytic leukemia (AMoL). Anti-human CD47 antibody, alone or combined with cytosine arabinoside (Ara-C), was used to treat the mice with AMoL for 7-14 days, and therapeutic efficacy was assessed. LSCs were cultured together with mouse macrophages in culture medium containing anti-CD47 or anti-CD45 monoclonal antibody for 2 hours, to observe the phagocytic ability of macrophages to LSCs. RESULTS: CD34+CD38- LSCs existed among THP-1 cells, with a content of about (0.12±0.06)%, and a mouse model of AML was successfully established after the purified CD34+CD38- LSCs (97.0%±1.7%) were transplanted into NOD/SCID mice. The in vivo experiment showed that mice with AMoL had the most significant decrease in CD33+CD45+ leukemia cells in peripheral blood and bone marrow and survived the longest after being treated with Ara-C (7 days) plus anti-CD47 monoclonal antibody (14 days) (P<0.01). After 2 hours of in vitro culture, the phagocytic index in the culture medium containing anti-CD47 monoclonal antibody was significantly higher than in the culture medium containing anti-CD45 monoclonal antibody (76.9%±12.2% vs 7.60%±2.4%; P<0.05). CONCLUSIONS: High expression of CD47 is an adverse prognostic factor in AML. Combination therapy with anti-CD47 monoclonal antibody and Ara-C can effectively eliminate leukemia cells and LSCs, demonstrating great clinical significance in curing AML.

导出引用

王颖超，冯磊，殷楚云，马丽娜，魏永纬，王春美，盛光耀. 抗CD47抗体联合阿糖胞苷靶向治疗NOD/SCID小鼠单核细胞白血病的研究[J]. 中国当代儿科杂志. 2013, 15(7): 577-582 https://doi.org/10.7499/j.issn.1008-8830.2013.07.017

WANG Ying-Chao, FENG Lei, YIN Chu-Yun, MA Li-Na, WEI Yong-Wei, WANG Chun-Mei, SHENG Guang-Yao. Effectiveness of Anti-CD47 antibody and Ara-C combination targeting therapy NOD/SCID mouse of myeloid leukemia[J]. Chinese Journal of Contemporary Pediatrics. 2013, 15(7): 577-582 https://doi.org/10.7499/j.issn.1008-8830.2013.07.017

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