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Blood, 1 December 2007, Vol. 110, No. 12, pp. 3842-3852. Prepublished online as a Blood First Edition Paper on August 23, 2007; DOI 10.1182/blood-2007-04-087346. This Article Full Text (PDF) Supplemental Table and Figures All Versions of this Article: blood-2007-04-087346v1 110/12/3842    most recent Alert me when this article is cited Alert me if a correction is posted 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 CrossRef Citing Articles via Google Scholar Google Scholar Articles by Deschamps, M. Articles by Ferrand, C. Search for Related Content PubMed PubMed Citation Articles by Deschamps, M. Articles by Ferrand, C. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted April 26, 2007 Accepted July 20, 2007 Deletions within the HSV-tk transgene in long-lasting circulating gene-modified T-cells infused with a hematopoietic graft Marina Deschamps, Patricia Mercier, Jean Marie Certoux, Carole Henry, Bruno Lioure, Celine Pagneux, Jean Yves Cahn, Eric Deconinck, Pierre Tiberghien, Eric Robinet, and Christophe Ferrand* INSERM, U645, Besancon, France Universite de Franche-Comte, IFR133, Besancon, France Immuno-Molecular Therapeutics Laboratory, EFS-BFC, Besancon, France Hematology Department, CHU Strasbourg, Strasbourg, France Clinical Biomonitoring Laboratory, EFS-BFC, Besancon, France INSERM U823, University Hospital Grenoble, Universite Joseph Fourier, Grenoble, France Hematology Department, CHU Besancon, Besancon, France * Corresponding author; email: christophe.ferrand{at}efs.sante.fr. In our previous phase I/II study aimed at controlling graft-versus-host disease, 12 patients received Herpes-simplex-virus-thymidine kinase (HSV-tk+) / Neomycine-Phosphotransferase (NeoR+)-expressing donor gene-modified T-cells (GMCs) and an HLA-identical sibling T-cell-depleted BMT. This study's objective was to follow-up, to quantify and characterize persistently circulating GMCs more than 10 years after BMT. Circulating GMCs remain detectable in all 4 evaluable patients. However, NeoR- and HSV-tk-PCR differently quantified in vivo counts, suggesting deletions within the HSV-tk gene. Further experiments, including a novel "transgene walking" PCR method, confirmed the presence of deletions. The deletions were unique, patient-specific, present in most circulating GMCs expressing NeoR and shown to occur at time of GMC production. Unique patient-specific retroviral insertion sites were found in all GMCs capable of in vitro expansion/cloning as well. These findings suggest a rare initial gene deletion event and an in vivo survival advantage of rare GMC clones resulting from an anti-HSV-tk immune response and/or Ganciclovir treatment. In conclusion, we show that donor mature T-cells infused with a T-cell depleted graft persist in vivo for more than a decade. These cells, containing transgene deletions and subjected to significant in vivo selection, represent a small fraction of T-cells infused at transplantation. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

	Copyright © 2007 by American Society of Hematology         Online ISSN: 1528-0020