Transfer of Drug-Resistant Genes Into Hemopoietic Cell Using Retroviral Vecto

为了建立一种适合的方法，将红细胞转移到红细胞干细胞使用逆转录病毒载体系统，我们固定了几种不同的逆转录病毒载体系统，命名为ZGX-neoGM，其中一种与人类颗粒细胞/宏观细胞克隆刺激因子cDNA在骨髓可移植性肉瘤病毒（MPSV）中集成了新霉素耐药性基因，pLGMRNL和pLGMSRNL是由murine leukemia病毒（MuLV）衍生的，并由新霉素耐药性基因和人类颗粒/宏观基因组成的。这些逆转录病毒载体在K562细胞中的传染性有效性，起源于人类风湿性白血病，ZGX-neoGM为0.7%，pLGMRNL为0.2，pLGMSRNL为0.1%，相应地。另一方面，受感染的K562细胞的GM-CSF生产低于ZGX-neoGM的3 pg/ml，pLGMRNL的160 pg/ml，pLGMSRNL的2800 pg/ml。GM-CSF生产受感染的K562细胞的活动，以使PLGMRNL能够在体外和体内发挥功能。However，这种逆转录病毒载体系统的感染效应是不够的，无法转移exogenous genes to bone marrow hematopoietic stem cells因为这些stem cells在bone marrow cells中存在0.1%。我们现在正在探索一个更有效的系统，以便使用逆转录病毒载体进行基因转移。

In order to establish a suitable method for gene transfer to hematopoietic stem cells using retroーviral vectors we constituted several different retro-viral vector systems, namely, ZGX-neoGM which was integrated with human granelocyte/ macrophage colony-stimulating factor cDNA in myelo-ptoliferative sarcoma virus (MPSV) containing neomycin-resistant gene, pLGMRNL and pLGMSRNL which were originated from murine leukemia viruses (MuLV) and constructed with neomysin-resistant gene and human granulocyte/macrophage gene. Infectious Efficacy of these retroviral vectors in K562 cells,which origenated from human erythroleukemia, was 0.7% by ZGX-neoGM, 0.2 by pLGMRNL and 0.1% by pLGMSRNL, respectively. On the other hand, GM-CSF production of infected K562 cells was below 3pg/ml by ZGX-neoGM, 160pg/ml by pLGMRNL and 2800pg/ml by pLGMSRNL. The GM-CSF producing activities of infected K562 cells with PLGMRNL appears to be enough to function in vitro and in vivo. However, infectious efficacy of this retroviral vector system is not enough to transfer exogenous genes to bone marrow hematopoietic stem cells because these stem cells exist at 0.1% in bone marrow cells. We are now exploring the more effective system for gene transfer using retroviral vectors.