Abstract
FOXP3+ regulatory T cells (Tregs) play an important role in controlling immune activation, maintenance of homeostasis and the prevention of autoimmunity. Much effort has been focused in assessing their potential defects in certain human diseases and in developing potential Treg immunotherapy to cure autoimmune conditions. One cardinal feature of Tregs is their ability to suppress the activation and proliferation of effector T cells. The in vitro suppression assay is a convenient and relatively reliable method to quantify their suppressive function and validate their identity as true Tregs. This protocol describes a reliable, nonradioactive in vitro suppression assay to assess the immunosuppressive property of human Tregs in inhibiting proliferation of dye-labeled responder T cells using non-irradiated, HLA-DR+ antigen presenting cells and soluble anti-CD3 antibodies as stimuli.
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References
Sakaguchi S, Miyara M, Costantino CM, Hafler DA (2010) FOXP3+ regulatory T cells in the human immune system. Nat Rev Immunol 10:490–500
Rudensky AY (2011) Regulatory T cells and Foxp3. Immunol Rev 241:260–8
van der Vliet HJ, Nieuwenhuis EE (2007) IPEX as a result of mutations in FOXP3. Clin Dev Immunol 2007:89017
Miyara M, Sakaguchi S (2011) Human FoxP3(+)CD4(+) regulatory T cells: their knowns and unknowns. Immunol Cell Biol 89:346–51
Cvetanovich GL, Hafler DA (2010) Human regulatory T cells in autoimmune diseases. Curr Opin Immunol 22:753–60
Sakaguchi S, Wing K, Onishi Y, Prieto-Martin P, Yamaguchi T (2009) Regulatory T cells: how do they suppress immune responses? Int Immunol 21:1105–11
Tran DQ, Glass DD, Uzel G, Darnell DA, Spalding C, Holland SM et al (2009) Analysis of adhesion molecules, target cells, and role of IL-2 in human FOXP3+ regulatory T cell suppressor function. J Immunol 182:2929–38
Tang Q, Adams JY, Tooley AJ, Bi M, Fife BT, Serra P et al (2006) Visualizing regulatory T cell control of autoimmune responses in nonobese diabetic mice. Nat Immunol 7:83–92
Onishi Y, Fehervari Z, Yamaguchi T, Sakaguchi S (2008) Foxp3+ natural regulatory T cells preferentially form aggregates on dendritic cells in vitro and actively inhibit their maturation. Proc Natl Acad Sci U S A 105:10113–10118
Quah BJC, Parish CR (2010) The use of carboxyfluorescein diacetate succinimidyl ester (CFSE) to monitor lymphocyte proliferation. J Vis Exp 44:e2259
Acknowledgement
This work was supported by institutional funding from the Department of Pediatrics, The University of Texas Medical School at Houston.
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Tran, D.Q. (2013). In Vitro Suppression Assay for Functional Assessment of Human Regulatory T Cells. In: Snow, A., Lenardo, M. (eds) Immune Homeostasis. Methods in Molecular Biology, vol 979. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-290-2_16
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DOI: https://doi.org/10.1007/978-1-62703-290-2_16
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-289-6
Online ISBN: 978-1-62703-290-2
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