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  

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

作　　者：王颖超[1] 冯磊[1] 殷楚云[1] 马丽娜[1] 魏永纬[1] 王春美[1] 盛光耀[1] 

机构地区：[1]郑州大学第一附属医院儿科,河南省高等学校临床医学重点学科开放实验室,河南郑州450052

出　　处：《中国当代儿科杂志》2013年第7期577-582,共6页Chinese Journal of Contemporary Pediatrics

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。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-CIM5 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 ~ CIM5 ~ 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.

关 键 词：急性髓细胞白血病 白血病干细胞 CD47 阿糖胞苷 靶向治疗 小鼠 

分 类 号：R733.7[医药卫生—肿瘤]