Abstract
Naturally occurring CD4+CD25+T regulatory cells originate from the thymus and play a central role regarding the maintenance of peripheral tolerance by suppression of autoreactive T-cell populations. However, T regulatory cells can have beneficial as well as harmful effects. On the one hand, they prevent a variety of autoimmune and inflammatory diseases; but on the other hand, they concomitantly inhibit antitumor immune reactions by suppressing tumor-specific T-cell responses. Therefore, these ambivalent properties of T regulatory cells require detailed investigation especially with respect to a potential therapeutic exploitation of these cells. A prerequisite for such analyses is the isolation of pure T regulatory cells and the establishment of functional tests for the analysis of their suppressive properties, since no specific markers for CD4+CD25+T regulatory cells are known so far. In this chapter, techniques applying immunomagnetic beads have been used to establish an efficient method for isolation of human as well as murine regulatory T-cells. A combination of positive and negative selection steps using immunomagnetic beads of different sizes yields preparations of functional, active CD4+CD25+regulatory T-cells with high purity. Additionally, co-culture assays for functional characterization of isolated CD4+CD25+regulatory T-cells are described that are able to detect their suppressive properties for conventional CD4+T-cells with high sensitivity.
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Jonuleit, H., Schmitt, E. (2005). Regulatory T-Cells in Antitumor Therapy Isolation and Functional Testing of CD4+CD25+Regulatory T-Cells. In: Ludewig, B., Hoffmann, M.W. (eds) Adoptive Immunotherapy: Methods and Protocols. Methods in Molecular Medicine™, vol 109. Humana Press. https://doi.org/10.1385/1-59259-862-5:285
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DOI: https://doi.org/10.1385/1-59259-862-5:285
Publisher Name: Humana Press
Print ISBN: 978-1-58829-406-7
Online ISBN: 978-1-59259-862-5
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