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Blood, 1 May 2004, Vol. 103, No. 9, pp. 3565-3572. Prepublished online as a Blood First Edition Paper on December 11, 2003; DOI 10.1182/blood-2003-09-3056. This Article Full Text (PDF) All Versions of this Article: 2003-09-3056v1 103/9/3565    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 HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Rauser, G. Articles by Topp, M. S Search for Related Content PubMed PubMed Citation Articles by Rauser, G. Articles by Topp, M. S Social Bookmarking                   What's this? Submitted September 5, 2003 Accepted December 3, 2003 Rapid generation of combined CMV-specific CD4+and CD8+ T-cell lines for adoptive transfer into allogeneic stem cell transplant recipients Georg Rauser, Hermann Einsele, Christian Sinzger, Dorothee Wernet, Gabriele Kuntz, Mario Assenmacher, John D M Campbell, and Max S Topp* Medizinische Klinik II, Universitaetsklinikum Tuebingen, Tuebingen, Germany Medizinische Virologie, Universitaetsklinikum Tuebingen, Tuebingen, Germany Transfusionsmedizin, Universitaetsklinikum Tuebingen, Tuebingen, Germany Miltenyi Biotec, Bergisch Gladbach, Germany * Corresponding author; email: max.topp{at}med.uni-tuebingen.de. Adoptive transfer of cytomegalovirus-specific T cells can restore long lasting virus-specific immunity and clear CMV-viremia in allogeneic stem cell transplant (SCT) recipients, if both CD4+ and CD8+ CMV-specific T-cells are detected in the recipient after transfer. Current protocols for generating virus-specific T cells utilize live virus, require leukapheresis of donor and remains time consuming. To circumvent these limitations, a clinical scale protocol was developed for generation of CMV-specific T cells by utilizing only autologous cellular and serum components derived from one single 500mL blood draw. CMV-specific T cells were stimulated simultaneously with CMV-specific MHC-I restricted peptides and CMV-antigen. Activated T cells were isolated with the interferon- secretion assay and expanded for 10 days. In 8 randomly selected CMV-seropositive donors on average 1.34x108 combined CD4+ and CD8+ CMV-specific T cells were generated as determined by antigen-triggered interferon- production. CMV-infected fibroblasts were efficiently lysed by the generated T cells and both CMV-specific CD4+ and CD8+ T-cells expanded if stimulated with natural processed antigen. On the other hand, CD4+ and CD8+ T-cell mediated alloreactivity of generated CMV-specific T-cell lines was reduced when compared to the starting population. In conclusion, the developed culture system allows rapid generation of allo-depleted and highly enriched combined CD4+ and CD8+ CMV-specific T cells under good manufactoring practice conditions. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: J. Diekmann, E. Adamopoulou, O. Beck, G. Rauser, S. Lurati, S. Tenzer, H. Einsele, H.-G. Rammensee, H. Schild, and M. S. Topp Processing of Two Latent Membrane Protein 1 MHC Class I Epitopes Requires Tripeptidyl Peptidase II Involvement J. Immunol., August 1, 2009; 183(3): 1587 - 1597. [Abstract] [Full Text] [PDF] K. Besold, N. Frankenberg, S. Pepperl-Klindworth, J. Kuball, M. Theobald, G. Hahn, and B. 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Locatelli Prospective simultaneous quantification of human cytomegalovirus-specific CD4+ and CD8+ T-cell reconstitution in young recipients of allogeneic hematopoietic stem cell transplants Blood, August 15, 2006; 108(4): 1406 - 1412. [Abstract] [Full Text] [PDF] O. Beck, M. S. Topp, U. Koehl, E. Roilides, M. Simitsopoulou, M. Hanisch, J. Sarfati, J. P. Latge, T. Klingebiel, H. Einsele, et al. Generation of highly purified and functionally active human TH1 cells against Aspergillus fumigatus Blood, March 15, 2006; 107(6): 2562 - 2569. [Abstract] [Full Text] [PDF] F. Aversa, A. Terenzi, A. Tabilio, F. Falzetti, A. Carotti, S. Ballanti, R. Felicini, F. Falcinelli, A. Velardi, L. Ruggeri, et al. Full Haplotype-Mismatched Hematopoietic Stem-Cell Transplantation: A Phase II Study in Patients With Acute Leukemia at High Risk of Relapse J. Clin. Oncol., May 20, 2005; 23(15): 3447 - 3454. [Abstract] [Full Text] [PDF]

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