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Blood, 1 July 2007, Vol. 110, No. 1, pp. 201-210. Prepublished online as a Blood First Edition Paper on March 19, 2007; DOI 10.1182/blood-2006-11-056168. This Article Full Text (PDF) All Versions of this Article: blood-2006-11-056168v1 110/1/201    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 Wolfl, M. Articles by Greenberg, P. D. Search for Related Content PubMed PubMed Citation Articles by Wolfl, M. Articles by Greenberg, P. D. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted November 8, 2006 Accepted March 2, 2007 Activation-induced expression of CD137 permits detection, isolation and expansion of the full repertoire of CD8+ T-cells responding to antigen without requiring knowledge of epitope-specificities Matthias Wolfl*, Jurgen Kuball, William Y. Ho, Hieu Nguyen, Thomas J. Manley, Marie Bleakley, and Philip D. Greenberg Program in Immunology, Fred Hutchinson Cancer Research Center, Seattle, WA, United States Department of Immunology, University of Washington School of Medicine, Seattle, WA, United States * Corresponding author; email: mwolfl{at}arcor.de. CD137 is a member of the TNFR-family with co-stimulatory function. Here we show that it also has many favorable characteristics as a surrogate marker for antigen-specific activation of human CD8+ T-cells. While undetectable on unstimulated CD8+ T-cells, it is uniformly upregulated 24h after stimulation on virtually all responding cells regardless of differentiation stage or profile of cytokine secretion, which circumvents limitations of current surrogate markers for defining the repertoire of responding cells based on only individual functions. Antibody-labeled responding CD137+ cells can be easily and efficiently isolated by flow-sorting or magnetic beads to substantially enrich antigen-specific T-cells. To test this approach for epitope discovery, we examined in vitro priming of naive T-cells from healthy donors to Wilms tumor antigen 1 (WT1), a protein over-expressed in various malignancies. Two overlapping pentadecamers were identified as immunogenic, and further analysis defined WT1(286-293) as the minimal amino acid sequence and HLA-Cw07 as the HLA restriction element. In conclusion, this approach appears to be an efficient and sensitive in vitro technique to rapidly identify and isolate antigen-specific CD8+ T-cells present at low frequencies and displaying heterogenous functional profiles, and does not require prior knowledge of the specific epitopes recognized or the HLA-restricting-elements. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: A. Machlenkin, R. Uzana, S. Frankenburg, G. Eisenberg, L. Eisenbach, J. Pitcovski, R. Gorodetsky, A. Nissan, T. Peretz, and M. Lotem Capture of Tumor Cell Membranes by Trogocytosis Facilitates Detection and Isolation of Tumor-Specific Functional CTLs Cancer Res., March 15, 2008; 68(6): 2006 - 2013. [Abstract] [Full Text] [PDF] S. R. Riddell Engineering Antitumor Immunity by T-Cell Adoptive Immunotherapy Hematology, January 1, 2007; 2007(1): 250 - 256. [Abstract] [Full Text] [PDF]

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