Abstract
The direct assessment of T helper (TH)-cell responses specific for antigens is essential to evaluate pathogenic and protective immunity. Presently, analysis and isolation of antigen-specific TH cells is restricted to cells that produce cytokines, or can be performed only with a rare selection of specific peptide major histocompatibility complex class II (MHC II) multimers. Here we report a new method that enables the assessment and isolation of TH cells specific for a defined antigen according to CD154 expression induced after stimulation in vitro. We show that antigen-induced CD154 expression is highly sensitive and specific for human and mouse antigen-specific TH cells. Moreover, the isolation of antigen-specific CD154+ TH cells necessitates only surface staining with antibodies, thereby enabling the fast generation of antigen-specific TH cell lines. Our approach allows assessment of TH cells with a defined specificity for the combined quantitative and qualitative analysis of TH-cell immunity as well as for the isolation of specific TH cells for targeted cellular immunotherapies.
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Acknowledgements
We thank A. Radbruch for critical discussion and U. Klein for critical reading of the manuscript. We thank Q. Phan, F. Kellndorfer and D. Huscher for technical help. This work was supported by the European Sixth Framework Program, grants LSHP-CT-2003-503240, LSHG-CT-2005-005203 and LSHB-CT-2004-512074, grants PTJ 0311597 and PTJ 0312102 (all grants were to A.T. and A.S.).
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Supplementary Fig. 1
Combined qualitative and quantitative assessment of Th cells specific for different antigens. (PDF 872 kb)
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Frentsch, M., Arbach, O., Kirchhoff, D. et al. Direct access to CD4+ T cells specific for defined antigens according to CD154 expression. Nat Med 11, 1118–1124 (2005). https://doi.org/10.1038/nm1292
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DOI: https://doi.org/10.1038/nm1292
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