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

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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.