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Updated: Apr 25, 2026

In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Naïve CD4+ T Cells Using a TGF-β-containing Protocol
Published on: December 30, 2016
Assessing the suppressive activity of Foxp3⁺ regulatory T cells
Christian Thomas Mayer1, Tim Sparwasser
1Institute of Infection Immunology, TWINCORE/Centre for Experimental and Clinical Infection Research; a joint venture between the Medical School Hannover (MHH) and the Helmholtz Centre for Infection Research (HZI), GmbH Feodor-Lynen-Str. 7, 30625, Hannover, Germany.
Abstract:
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.
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