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This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Chinnasamy, D. Articles by Candotti, F. Search for Related Content PubMed PubMed Citation Articles by Chinnasamy, D. Articles by Candotti, F. Related Collections Immunobiology Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 15 August 2000, Vol. 96, No. 4, pp. 1309-1316 GENE THERAPY Lentiviral-mediated gene transfer into human lymphocytes: role of HIV-1 accessory proteins Dhanalakshmi Chinnasamy, Nachimuthu Chinnasamy, Melissa J. Enriquez, Makoto Otsu, Richard A. Morgan, and Fabio Candotti From the Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD. Resting lymphocytes are refractory to gene transfer using Moloney murine leukemia virus (MMLV)-based retroviral vectors because of their quiescent status. Recently, it has been shown that lentiviral vectors are capable of transferring genes into nondividing and terminally differentiated cells. We used human immunodeficiency virus type-1 (HIV-1)-based vectors expressing enhanced green fluorescent protein (EGFP) driven by different promoters (CMV, MPSV, or PGK) and investigated their ability to transduce human T- and B-cell lines, as well as resting or activated primary peripheral and umbilical cord blood lymphocytes. The effects of the presence or the absence of HIV-1 accessory proteins (Vif, Vpr, Vpu, and Nef) in the vector system were also assessed. Flow cytometry analysis showed no differences in the ability of these vectors of transferring the reporter gene into lymphocytic lines and mitogen-stimulated primary lymphocytes in the presence or the absence of HIV-1 accessory proteins (APs). Similarly, viral supernatants generated in the presence of accessory genes could efficiently transduce various subsets of resting lymphocytes and provide long-term expression of the transgene. No significant transduction-induced changes in cell activation or cycling status were observed and Alu-HIV-1 long terminal repeat polymerase chain reaction (LTR PCR) analysis demonstrated integration of the vector sequences at the molecular level. In contrast, in the absence of HIV-1 APs, lentiviral vectors failed to integrate and express the transgene in resting lymphocytes. These results show that transduction of primary resting lymphocytes with HIV-1-based vectors requires the presence of viral accessory proteins. © 2000 by The American Society of Hematology.   CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: X. Huang, A. C. Wilber, L. Bao, D. Tuong, J. Tolar, P. J. Orchard, B. L. Levine, C. H. June, R. S. McIvor, B. R. Blazar, et al. Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system Blood, January 15, 2006; 107(2): 483 - 491. [Abstract] [Full Text] [PDF] M. Lai, E. S. Zimmerman, V. Planelles, and J. Chen Activation of the ATR Pathway by Human Immunodeficiency Virus Type 1 Vpr Involves Its Direct Binding to Chromatin In Vivo J. Virol., December 15, 2005; 79(24): 15443 - 15451. [Abstract] [Full Text] [PDF] K. Shibuya, J. Shirakawa, T. Kameyama, S.-i. Honda, S. Tahara-Hanaoka, A. Miyamoto, M. Onodera, T. Sumida, H. 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