Generation of activated and antigen-specific T cells with cytotoxic activity after co-culture with dendritic cells |
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Authors: | Angela Märten Tim Greten Carsten Ziske Sabine Renoth Björn Schöttker Peter Buttgereit Frank Schakowski Alexander von Rücker Tilman Sauerbruch Ingo G Schmidt-Wolf |
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Institution: | Department of Internal Medicine I, Rheinische Friedrich-Wilhelms-University, Bonn, Germany. |
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Abstract: | Co-culturing of immunological effector cells with antigen-pulsed DC leads to an increase of cytotoxic activity against antigen-expressing tumour cells. Using this approach, we could detect up to 2.8% antigen-specific CTLs after co-culture with antigen-pulsed DC. However, the required high effector cell numbers remain a major obstacle in immunotherapy. In this study, we show an approach for generating activated and antigen-specific effector cells that enables us to decrease effector to target cell ratios. We used an interferon-gamma secretion assay to enrich activated effector cells after co-culture with antigen-pulsed dendritic cells (DC). Purified immunological effector cells lysed 58.3% of antigen-expressing tumour cells at an effector to target ratio of 1:1. Furthermore, using MHC-IgG complexes, we enriched effector cells expressing antigen-specific T-cell receptor after co-culture with DC. Performing ELISpot, flow cytometry and TCR analysis, we could show a significant increase of activated and specific TCR-expressing effector cells after co-culture with DC. |
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