title: The purification and functional analysis of human CD4+CD25high regulatory T cells.
abstract: Regulatory T cells were initially identified and isolated in the mouse, by virtue of their endogenous expression of CD25 (IL-2R alphachain) and shown to inhibit both the in vivo development of autoimmunity and the in vitro proliferation of nonregulatory, CD4+CD25- T cells. In contrast to mouse cells, human regulatory T cells are not purified by isolating all CD25-expressing CD4 T cells ex vivo. Such cells can be isolated by targeting only the small percentage of human CD4 T cells that express high levels of CD25. This is best achieved by FACS sorting using the level of CD25 expressed on CD4- T cells to place the gate for discriminating high expression of CD25. This unit provides two widely used methods to isolate (FACS) or to enrich (magnetic beads) human CD4+CD25+ regulatory T cells from blood, along with an in vitro coculture assay to measure the anergic and suppressive features of human CD4+CD25+ regulatory T cells.
authors.affiliation: Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.
authors.forename: Clare M
authors.initials: CM
authors.lastname: Baecher-Allan
authors.affiliation: Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.
authors.forename: David A
authors.initials: DA
authors.lastname: Hafler
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000502:physiology*
chemical_list: D053645:Interleukin-2 Receptor alpha Subunit