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Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector

Gene Therapy volume 10, pages 1446–1457 (2003)Cite this article

Abstract

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.

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Acknowledgements

We thank Dr IM Verma (Salk Institute, La Jolla, CA, USA) and Cell Genesys for providing HIV vector constructs. We also express our gratitude to Drs DR Littman, T Kitamura, K Ohyashiki, F Nagamura (University of Tokyo), and G Veres (Cell Genesys) for kindly providing us with pSV-Ψ−-env−-MLV plasmid, pMX plasmid, OM9; 22 cells, IMS-M2 cells and helpful discussion, respectively. We are grateful to Dr H Urano (Ikuryo clinic, Tokyo, Japan) for providing us with cord blood samples. We also appreciate Dr T Iseki and the hospital staff of the Research Hospital of Institute of Medical Science, University of Tokyo for their support in sample collection from patients. We are indebted to Drs N Watanabe and D Sugiyama (University of Tokyo) for suggestions regarding the multicolor FCM and to Dr Y Nakazaki, Dr E Sasaki, Mr K Takahashi, Mrs M Oiwa, and Mrs S Suzuki (University of Tokyo) for technical support. YB and YS are awardees of Research Fellowship of the Japan Society of the Promotion of Science for Young Scientists. This work was supported in part by a Grant-in-aid from the Ministry of Education, Science, Sports and Culture, and the Ministry of Health, Labor and Welfare of Japan.

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Author notes

  1. Y Bai & H Miyoshi

    Present address: Technology and Development Team for Biosignal Program, Bioresource Center, The Institute of Physical and Chemical Research, Tsakuba, Japan

  2. Y Bai and Y Soda: The first two authors equally contributed to this work

Authors and Affiliations

  1. Division of Molecular Therapy, Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Y Bai, Y Soda, K Izawa, T Tanabe, X Kang, A Tojo, S Asano & K Tani

  2. Department of Virology and Preventive Medicine, Gunma University School of Medicine, Maebashi, Japan

    Y Soda & H Hoshino

  3. Age Dimension Team, Age Dimension Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan

    T Tanabe

  4. Department of Immunology, Institute of Basic Medical Science, University of Tsukuba, Tsukuba, Japan

    H Miyoshi

  5. Division of Molecular and Clinical Genetics, Department of Molecular Genetics, Medical Institute of Bioregulation, Kyushu University, Beppu, Japan

    K Tani

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Bai, Y., Soda, Y., Izawa, K. et al. Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector. Gene Ther 10, 1446–1457 (2003). https://doi.org/10.1038/sj.gt.3302026

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