Co-expression of TNFR2 and CD25 identifies more of the functional CD4+FOXP3+ regulatory T cells in human peripheral blood

Eur J Immunol. 2010 Apr;40(4):1099-106. doi: 10.1002/eji.200940022.


Previously, we found that co-expression of CD25 and TNFR2 identified the most suppressive subset of mouse Treg. In this study, we report that human peripheral blood (PB) FOXP3(+) cells present in CD25(high), CD25(low) and even CD25(-) subsets of CD4(+) cells expressed high levels of TNFR2. Consequently, TNFR2-expressing CD4(+)CD25(+) Treg included all of the FOXP3(+) cells present in the CD4(+)CD25(high) subset as well as a substantial proportion of the FOXP3(+) cells present in the CD4(+)CD25(low) subset. Flow cytometric analysis of PB identified five-fold more Treg, determined by FOXP3 expression, in the CD4(+)CD25(+)TNFR2(+) subset than in the CD4(+)CD25(high) subset. In addition, similar levels of FOXP3(+) cells were identified in both the CD4(+)CD25(+)TNFR2(+) and CD4(+)CD25(+)CD127(low/-) subsets. Furthermore, the CD4(+)CD25(+)TNFR2(+) subset expressed high levels of CTLA-4, CD45RO, CCR4 and low levels of CD45RA and CD127, a phenotype characteristic of Treg. Upon TCR stimulation, human PB CD4(+)CD25(+)TNFR2(+) cells were anergic and markedly inhibited the proliferation and cytokine production of co-cultured T-responder cells. In contrast, CD4(+)CD25(+)TNFR2(-) and CD4(+)CD25(-)TNFR2(+) T cells did not show inhibitory activity. As some non-Treg express TNFR2, the combination of CD25 and TNFR2 must be used to identify a larger population of human Treg, a population that may prove to be of diagnostic and therapeutic benefit in cancer and autoimmune diseases.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Adult
  • Antigen Presentation
  • Antigens, CD / analysis
  • CTLA-4 Antigen
  • Cells, Cultured / immunology
  • Cells, Cultured / metabolism
  • Coculture Techniques
  • Flow Cytometry
  • Forkhead Transcription Factors / analysis*
  • Humans
  • Immunophenotyping
  • Interferon-gamma / biosynthesis
  • Interleukin-2 Receptor alpha Subunit / analysis
  • Interleukin-2 Receptor alpha Subunit / biosynthesis*
  • Interleukin-7 Receptor alpha Subunit / analysis
  • Leukocyte Common Antigens / analysis
  • Lymphocyte Activation / drug effects
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Tumor Necrosis Factor, Type II / analysis
  • Receptors, Tumor Necrosis Factor, Type II / biosynthesis*
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / metabolism
  • T-Lymphocytes, Regulatory / classification*
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / metabolism


  • Antigens, CD
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • Ctla4 protein, mouse
  • FOXP3 protein, human
  • Forkhead Transcription Factors
  • IL2RA protein, human
  • Interleukin-2 Receptor alpha Subunit
  • Interleukin-7 Receptor alpha Subunit
  • Receptors, Antigen, T-Cell
  • Receptors, Tumor Necrosis Factor, Type II
  • Interferon-gamma
  • Leukocyte Common Antigens