Abstract
Although quantitative identification and viable enrichment of natural regulatory T cells (T-regs) in humans are problematic, such steps would greatly facilitate the analysis of these cells in disease states. In an attempt to identify markers that are sensitive and specific for human T-regs, we analyzed the expression of fourteen intracellular and cell surface markers on human CD4+ cells. Many markers were partially selective for CD25hi T-regs, but consistent and specific discrimination of functional T-regs was only made possible by focus on CD127, the alpha chain of the IL-7 receptor. Although most CD4+ human T cells express CD127, T-regs exhibiting suppressive activity in vitro display distinctly lower surface expression of this marker, irrespective of their level of CD25 expression. Sorted cells with the surface phenotype CD4+CD25+CD127low had higher levels of intracellular FOXP3 and CTLA-4 and, as determined by functional assays, were suppressive, hypoproliferative, and poorly responsive to TCR signaling. The CD4+CD25+CD127low phenotype was also found to be characteristic of T-regs found in mice and in rhesus macaques. This surface phenotype should allow for quantitative studies of regulatory T cells in disease states as well as for enrichment of live regulatory T cells for functional analyses and/or expansion in vitro.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 41-52 |
| Number of pages | 12 |
| Journal | Journal of Immunological Methods |
| Volume | 319 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jan 30 2007 |
| Externally published | Yes |
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Keywords
- Cell surface molecules
- Human
- T cells
- Transcription factors
ASJC Scopus subject areas
- Biotechnology
- Immunology
Cite this
Human CD4+ regulatory T cells express lower levels of the IL-7 receptor alpha chain (CD127), allowing consistent identification and sorting of live cells. / Hartigan-O'Connor, Dennis; Poon, Chungkee; Sinclair, Elizabeth; McCune, Joseph M.
In: Journal of Immunological Methods, Vol. 319, No. 1-2, 30.01.2007, p. 41-52.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Human CD4+ regulatory T cells express lower levels of the IL-7 receptor alpha chain (CD127), allowing consistent identification and sorting of live cells
AU - Hartigan-O'Connor, Dennis
AU - Poon, Chungkee
AU - Sinclair, Elizabeth
AU - McCune, Joseph M.
PY - 2007/1/30
Y1 - 2007/1/30
N2 - Although quantitative identification and viable enrichment of natural regulatory T cells (T-regs) in humans are problematic, such steps would greatly facilitate the analysis of these cells in disease states. In an attempt to identify markers that are sensitive and specific for human T-regs, we analyzed the expression of fourteen intracellular and cell surface markers on human CD4+ cells. Many markers were partially selective for CD25hi T-regs, but consistent and specific discrimination of functional T-regs was only made possible by focus on CD127, the alpha chain of the IL-7 receptor. Although most CD4+ human T cells express CD127, T-regs exhibiting suppressive activity in vitro display distinctly lower surface expression of this marker, irrespective of their level of CD25 expression. Sorted cells with the surface phenotype CD4+CD25+CD127low had higher levels of intracellular FOXP3 and CTLA-4 and, as determined by functional assays, were suppressive, hypoproliferative, and poorly responsive to TCR signaling. The CD4+CD25+CD127low phenotype was also found to be characteristic of T-regs found in mice and in rhesus macaques. This surface phenotype should allow for quantitative studies of regulatory T cells in disease states as well as for enrichment of live regulatory T cells for functional analyses and/or expansion in vitro.
AB - Although quantitative identification and viable enrichment of natural regulatory T cells (T-regs) in humans are problematic, such steps would greatly facilitate the analysis of these cells in disease states. In an attempt to identify markers that are sensitive and specific for human T-regs, we analyzed the expression of fourteen intracellular and cell surface markers on human CD4+ cells. Many markers were partially selective for CD25hi T-regs, but consistent and specific discrimination of functional T-regs was only made possible by focus on CD127, the alpha chain of the IL-7 receptor. Although most CD4+ human T cells express CD127, T-regs exhibiting suppressive activity in vitro display distinctly lower surface expression of this marker, irrespective of their level of CD25 expression. Sorted cells with the surface phenotype CD4+CD25+CD127low had higher levels of intracellular FOXP3 and CTLA-4 and, as determined by functional assays, were suppressive, hypoproliferative, and poorly responsive to TCR signaling. The CD4+CD25+CD127low phenotype was also found to be characteristic of T-regs found in mice and in rhesus macaques. This surface phenotype should allow for quantitative studies of regulatory T cells in disease states as well as for enrichment of live regulatory T cells for functional analyses and/or expansion in vitro.
KW - Cell surface molecules
KW - Human
KW - T cells
KW - Transcription factors
UR - http://www.scopus.com/inward/record.url?scp=33846283803&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33846283803&partnerID=8YFLogxK
U2 - 10.1016/j.jim.2006.10.008
DO - 10.1016/j.jim.2006.10.008
M3 - Article
C2 - 17173927
AN - SCOPUS:33846283803
VL - 319
SP - 41
EP - 52
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -