裸鼠腹腔输注体外扩增的阵发性睡眠性血红蛋白尿症病人骨髓CD34+CD59+细胞的研究

应用BALB／c裸鼠为移植模型，将在体外扩增的阵发性睡眠性血红蛋白尿症（paroxysmal nocturnal hemoglobinuria，PNH）病人的CD34^＋CD59^＋细胞进行移植。研究其增殖能力和重建造血的情况，为实现PNH患者进行临床自体骨髓移植（ABMT）或自体外周血干细胞移植（APBSCT）奠定基础。本研究利用免疫磁珠双阳性分选法。从PNH患者骨髓中分离出足够数量的CD34^＋CD59^＋细胞进行体外扩增，并将体外扩增的细胞输注给接受亚致死剂量照射的BALB／c裸鼠。结果显示：移植6周后，用流式细胞术和DNA检测方法均检测到裸鼠骨髓、脾脏和外周血中人的CD45^＋细胞，而接受PNH病人CD34^＋CD59^＋细胞移植后的裸鼠，其血常规指标有一定恢复，但并未完全恢复。结论：PNH病人骨髓CD34^＋CD59^＋细胞体外扩增后仍能保持造血祖细胞的生物特性，在照射的裸鼠中能重建造血。

Nude Mice Intraperitoneally Transfused with Ex Vivo Expanded Bone Marrow CD34 + CD59 + Cells from Patients with Paroxysmal Nocturnal Hemoglobinuria

Ex vivo expanded human bone marrow CD34~+CD59~+ cells from patients with paroxysmal nocturnal hemoglobinuria (PNH) were transplanted into BALB/c mice in order to investigate their proliferation ability and reconstruction of hemopoiesis, and to lay the groundwork for clinical ABMT/APBSCT in PNH patients. CD34~+CD59~+ cells were selected from the bone marrow mononuclear cells in PNH patients by using immunomagnetic positive double sorting. Sublethally irradiated BALB/c mice were transplanted with CD34~+CD59~+ cells enriched from bone narrow of PNH patients.The results showed that human CD45~+ cells were detected in the bone marrow, spleen and peripheral blood of the nude mice by flow cytometry and DNA analysis at 6 weeks post-transplant. Blood routine indicators of nude mice were found to recover to some extent, but did not fully recover. It is concluded that ex vivo expanded bone marrow CD34~+CD59~+ cells from patients with paroxysmal nocturnal hemoglobinuria could keep their biological characteristics and ability to reconstruct hemopoiesis in irradiated BALB/c mice.