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Prepublished online as a Blood First Edition Paper on July 5, 2002; DOI 10.1182/blood-2002-02-0599. This Article Full Text Full Text (PDF) All Versions of this Article: 2002-02-0599v1 100/4/1257    most recent 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 Nicolini, F. E. Articles by Eaves, C. J. Search for Related Content PubMed PubMed Citation Articles by Nicolini, F. E. Articles by Eaves, C. J. Related Collections Hematopoiesis and Stem Cells Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 15 August 2002, Vol. 100, No. 4, pp. 1257-1264 GENE THERAPY Expression of a human -globin transgene in erythroid cells derived from retrovirally transduced transplantable human fetal liver and cord blood cells Franck E. Nicolini, Suzan Imren, Il-Hoan Oh, R. Keith Humphries, Philippe Leboulch, Mary E. Fabry, Ronald L. Nagel, and Connie J. Eaves From the Terry Fox Laboratory, British Columbia Cancer Agency and University of British Columbia, Vancouver, BC, Canada; Harvard-MIT, Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge; Genetix Pharmaceuticals, Cambridge, MA; and Albert Einstein College of Medicine, Bronx, NY. Transfer of therapeutic genes to human hematopoietic stem cells (HSCs) using complex vectors at clinically relevant efficiencies remains a major challenge. Recently we described a stable retroviral vector that sustains long-term expression of green fluorescent protein (GFP) and a human -globin gene in the erythroid progeny of transduced murine HSCs. We now report the efficient transduction of primitive human CD34+ fetal liver or cord blood cells with this vector and expression of the -globin transgene in the erythroid progeny of these human cells for at least 2 months. After growth factor prestimulation and then a 2- to 3-day exposure to the virus, 35% to 55% GFP+ progeny were seen in assays of transduced colony-forming cells, primitive erythroid precursors that generate large numbers of glycophorin A+ cells in 3-week suspension cultures, and 6-week long-term culture-initiating cells. In immunodeficient mice injected with unselected infected cells, 5% to 15% of the human cells regenerated in the marrow (including the erythroid cells) were GFP+ 3 and 6 weeks after transplantation. Importantly, the numbers of GFP+ human lymphoid and either granulopoietic or erythroid cells in individual mice 6 weeks after transplantation were significantly correlated, indicative of the initial transduction of human multipotent cells with in vivo repopulating activity. Expression of the transduced -globin gene in human cells obtained directly from the mice or after their differentiation into erythroid cells in vitro was demonstrated by reverse transcriptase-polymerase chain reaction using specific primers. These experiments represent a significant step toward the realization of a gene therapy approach for human -globin gene disorders. © 2002 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: G. F. Atweh, J. DeSimone, Y. Saunthararajah, H. Fathallah, R. S. Weinberg, R. L. Nagel, M. E. Fabry, and R. J. Adams Hemoglobinopathies Hematology, January 1, 2003; 2003(1): 14 - 39. [Abstract] [Full Text] [PDF]

	Copyright © 2002 by American Society of Hematology         Online ISSN: 1528-0020