Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector
Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector
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DOI：
10.1038/sj.gt.3302026
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发表时间：
2003-08-01
期刊：
影响因子：
5.1
通讯作者：
Tani, K
中科院分区：
文献类型：
--
作者：
Bai, Y;Soda, Y;Tani, K
人血细胞（包括造血干细胞）的基因转导的困难阻碍了血液疾病的基因治疗应用的发展，鼓励了新基因递送系统的开发和使用。在这项研究中，我们使用了第三代自失活（SIN）慢病毒载体系统的基础上，人类免疫缺陷病毒1型（HIV-1），以提高转导效率和防止载体相关的毒性。在人白血病细胞系中直接比较基于HIV-1的载体与莫洛尼鼠白血病病毒（MLV）SIN载体的转导效率。HIV的初始转导效率几乎为100%，而MLV载体的初始转导效率低于50%。在从白血病或骨髓瘤患者获得的11种类型的原代细胞中观察到类似的结果。在用HIV载体转导的细胞中，转基因表达持续8周，但用MLV载体则下降。此外，静息外周血淋巴细胞和CD 34（+）造血细胞成功地转导了HIV载体，但没有与MLV载体。最后，我们证实了载体基因整合在几乎所有的集落形成细胞转导的HIV载体，但不是与MLV载体。总之，这种慢病毒载体是一个很好的人血细胞基因转导系统，因为它的基因转导和宿主染色体整合效率高。
Difficulty in gene transduction of human blood cells, including hematopoietic stem cells, has hampered the development of gene therapy applications for hematological disorders, encouraging the development and use of new gene delivery systems. In this study, we used a third-generation self-inactivating (SIN) lentiviral vector system based on human immunodeficiency virus type 1 (HIV-1) to improve transduction efficiency and prevent vector-related toxicity. The transduction efficiency of the HIV-1-based vector was compared directly with the Moloney murine leukemia virus (MLV) SIN vector in human leukemia cell lines. Initial transduction efficiencies were almost 100% for the HIV and less than 50% for the MLV vectors. Similar results were observed in 11 types of primary cells obtained from leukemia or myeloma patients. Transgene expression persisted for 8 weeks in cells transduced with the HIV vector, but declined with the MLV vector. In addition, resting peripheral blood lymphocytes and CD34(+) hematopoietic cells were transduced successfully with the HIV vector, but not with the MLV vector. Finally, we confirmed vector gene integration in almost all colony-forming cells transduced with the HIV vector, but not with the MLV vector. In conclusion, this lentiviral vector is an excellent gene transduction system for human blood cells because of its high gene transduction and host chromosome integration efficiency.