title: Flow cytometric detection of human regulatory T cells.
abstract: Tregs are absolutely required for the maintenance of self tolerance in mouse and man. Major abnormalities in Treg number or function cause rare but fatal syndromes with autoimmune, allergic and inflammatory features. Whether subtle Treg abnormalities contribute to the pathogenesis of sporadic autoimmune, allergic and immunoinflammatory diseases in man remains controversial. Robust methods for identifying and isolating human Tregs in patients and healthy controls are essential if we are to understand their role in these increasingly common diseases. We have outlined below a flow cytometric technique to detect and isolate the entire human Treg population based on expression of CD4, CD25, and CD127. Use of a number of additional antibodies for defining subsets within the Treg compartment is described. For analysis, anti-Foxp3 can be added to the cocktail, but the necessity for fixation and permeabilisation may reduce the signal from other antibodies.
authors.affiliation: T Cell Biology Research Program, Centenary Institute and Faculty of Medicine, University of Sydney, Sydney, NSW, Australia.
authors.forename: Barbara
authors.initials: B
authors.lastname: Fazekas de St Groth
authors.forename: Erhua
authors.initials: E
authors.lastname: Zhu
authors.forename: Suzanne
authors.initials: S
authors.lastname: Asad
authors.forename: Loretta
authors.initials: L
authors.lastname: Lee
mesh_terms: D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods*; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D050378:T-Lymphocytes, Regulatory / Q000166:cytology*
chemical_list: C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors