Immunophenotyping of Human Regulatory T Cells

Regulatory T cells, also known as Tregs, play a pivotal role in maintaining homeostasis of the immune system and self-tolerance. Tregs express CD3, CD4, CD25, and FOXP3 but lack CD127. CD4 and CD3 identify helper T lymphocytes, of which Tregs are a subset. CD25 is IL-2Rα, an essential activation mar...

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Veröffentlicht in:Methods in molecular biology (Clifton, N.J.) Jg. 2032; S. 141
1. Verfasser: Staats, Janet
Format: Journal Article
Sprache:Englisch
Veröffentlicht: United States 2019
Schlagworte:
CD3 Complex - immunology
CD4 Antigens - immunology
Flow Cytometry - methods
Forkhead Transcription Factors - immunology
Humans
Immunophenotyping - methods
Interleukin-2 Receptor alpha Subunit - immunology
Interleukin-7 Receptor alpha Subunit - immunology
Receptors, Interleukin-7 - immunology
T-Lymphocytes, Regulatory - immunology
Suppressor T cells
Flow cytometry
Effector tregs
Tregs
Immunophenotyping
CD127
iTregs
FoxP3
nTregs
ISSN:1940-6029, 1940-6029
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Zusammenfassung:Regulatory T cells, also known as Tregs, play a pivotal role in maintaining homeostasis of the immune system and self-tolerance. Tregs express CD3, CD4, CD25, and FOXP3 but lack CD127. CD4 and CD3 identify helper T lymphocytes, of which Tregs are a subset. CD25 is IL-2Rα, an essential activation marker that is expressed in high levels on Tregs. FOXP3 is the canonical transcription factor, important in the development, maintenance, and identification of Tregs. CD127 is IL-7 receptor, expressed inversely with suppression, and is therefore downregulated on Tregs. Flow cytometry is a powerful tool that is capable of simultaneously measuring Tregs along with several markers associated with subpopulations of Tregs, activation, maturation, proliferation, and surrogates of functional suppression. This chapter describes a multicolor flow cytometry-based approach to measure human Tregs, including details for surface staining, fixation/permeabilization, intracellular/intranuclear staining, acquisition of samples on a flow cytometer, plus analysis and interpretation of resulting FCS files.
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ISSN:1940-6029
1940-6029
DOI:10.1007/978-1-4939-9650-6_9