Assessing the Suppressive Activity of Foxp3+ Regulatory T Cells
Foxp3+ regulatory T cells (Tregs) balance the mammalian immune system by mechanisms that are yet to be elucidated in their entirety. Methods employed to quantify the regulatory activity of Tregs in vitro are an important tool in cellular immunology, but can be technically demanding and subjected to variation. In this manuscript, we describe in detail a robust Treg suppression assay based on the flow cytometric quantification of both CD4+ and CD8+ effector T cell functions. This method can provide valuable insights into the immunosuppressive activity of Foxp3+ Tregs and is versatile with regard to genetic or pharmacologic manipulations. Additionally, novel regulatory immune cells can be characterized by using this assay.
Key wordsFoxp3 Treg Suppression assay Tolerance Autoimmunity Cancer
We thank Maxine Swallow, Friederike Kruse, Melanie Gohmert, Martina Thiele, and Maike Hegemann for expert technical assistance and Dr. Katharina Lahl, Catharina Arnold-Schrauf, Christina Hesse, and Venkateswaran Ganesh for critical reading of the manuscript. C.T.M. was supported by the German National Academic Foundation. We would further like to thank the Cell Sorting Core Facility of the Hannover Medical School supported in part by the Braukmann-Wittenberg-Herz-Stiftung and Deutsche Forschungsgemeinschaft.
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