UNIT 7.4B The Purification and Functional Analysis of Human CD4+CD25high Regulatory T Cells
Published Online: 1 MAY 2006
Copyright © 2006 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Baecher-Allan, C. M. and Hafler, D. A. 2006. The Purification and Functional Analysis of Human CD4+CD25high Regulatory T Cells. Current Protocols in Immunology. 72:7.4B.1–7.4B.12.
- Published Online: 1 MAY 2006
- Published Print: APR 2006
Regulatory T cells were initially identified and isolated in the mouse, by virtue of their endogenous expression of CD25 (IL-2R αchain) 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.
- T cell subpopulation;