Activation requirements for the induction of CD4+CD25+ T cell suppressor function

Eur J Immunol. 2004 Feb;34(2):366-76. doi: 10.1002/eji.200324455.


The in vivo differentiation/survival of CD4(+)CD25(+) T suppressor cells is dependent on IL-2 and CD28-mediated costimulatory signals. To determine the cytokine and costimulatory requirements for CD25(+) T cells in vitro, we established a two-stage culture system where CD25(+) T cells were activated in a primary culture. In the subsequent culture, activated CD4(+)CD25(+) cells were then mixed with responders in order to assess their suppressor function. Pre-culture of CD25(+) T cells with anti-CD3 alone resulted in poor survival and minimal induction of suppressor activity. Pre-culture in the presence of anti-CD3 and IL-2 or IL-4, but not IL-6, IL-7, IL-9, IL-10 or IL-15, resulted in proliferation of the CD25(+) cells and induction of potent suppressor function. Inhibition of the interaction of CD28 or cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) with CD80/CD86 in the pre-culture of CD4(+)CD25(+) cells did not prevent the induction of suppressor function. Furthermore, the inhibition of costimulatory signals did not inhibit the ability of fresh CD25(+) T cells to inhibit CD8(+) responders under conditions where activation of the responders was independent of CD80/CD86. These studies support the view that activation of CD25(+) T cells requires IL-2/IL-4 for their survival/differentiation into effector cells, but is independent of CD28/CTLA-4-mediated costimulation.

MeSH terms

  • Animals
  • Antigens, CD
  • Antigens, Differentiation / immunology
  • CD28 Antigens / immunology
  • CD4 Antigens / immunology*
  • CTLA-4 Antigen
  • Cell Division / immunology
  • Clonal Anergy
  • Female
  • Interleukin-2 / immunology
  • Lymphocyte Activation / immunology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Knockout
  • Mice, Transgenic
  • Receptors, Interleukin-2 / immunology*
  • T-Lymphocytes, Regulatory / cytology
  • T-Lymphocytes, Regulatory / immunology*


  • Antigens, CD
  • Antigens, Differentiation
  • CD28 Antigens
  • CD4 Antigens
  • CTLA-4 Antigen
  • Ctla4 protein, mouse
  • Interleukin-2
  • Receptors, Interleukin-2