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Infectious tolerance: human CD25(+) regulatory T cells convey suppressor activity to conventional CD4(+) T helper cells.

Jonuleit H, Schmitt E, Kakirman H, Stassen M, Knop J, Enk AH - J. Exp. Med. (2002)

Bottom Line: Regulatory CD4(+)CD25(+) T cells (Treg) are mandatory for maintaining immunologic self-tolerance.Coactivation of CD25(+) Treg cells with Treg cell-depleted CD4(+) T cells results in anergized CD4(+) T cells that in turn inhibit the activation of conventional, freshly isolated CD4(+) T helper (Th) cells.This explains previously published conflicting data on the role of TGF-beta in CD25(+) Treg cell-induced immunosuppression.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Mainz, 55101 Mainz, Germany. jonuleit@hautklinik.klinik.uni-mainz.de

ABSTRACT
Regulatory CD4(+)CD25(+) T cells (Treg) are mandatory for maintaining immunologic self-tolerance. We demonstrate that the cell-cell contact-mediated suppression of conventional CD4(+) T cells by human CD25(+) Treg cells is fixation resistant, independent from membrane-bound TGF-beta but requires activation and protein synthesis of CD25(+) Treg cells. Coactivation of CD25(+) Treg cells with Treg cell-depleted CD4(+) T cells results in anergized CD4(+) T cells that in turn inhibit the activation of conventional, freshly isolated CD4(+) T helper (Th) cells. This infectious suppressive activity, transferred from CD25(+) Treg cells via cell contact, is cell contact-independent and partially mediated by soluble transforming growth factor (TGF)-beta. The induction of suppressive properties in conventional CD4(+) Th cells represents a mechanism underlying the phenomenon of infectious tolerance. This explains previously published conflicting data on the role of TGF-beta in CD25(+) Treg cell-induced immunosuppression.

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The suppressive activity of human CD25+ Treg cells is independent from membrane-bound TGF-β. CD4+ and CD4+CD25+ T cells were isolated from buffy coats of healthy volunteers by positive selection using paramagnetic beads. (A) Surface expression of TGF-β by freshly isolated CD4+ T cells and CD25+ Treg cells in comparison to the same T cell populations preactivated for 48 h with anti-CD3 (OKT3, 1 μg/ml) and anti-CD28 mAb (CD28.2, 2 μg/ml). The figure shows the expression of TGF-β (LAP-biotinylated) detected by streptavidin-Cy5 and CD4 (RPAT4-FITC). (B) CD4+ T cells (105 cells per well) or CD25+ Treg cells (105 cells per well), alone or in coculture (1:1), were stimulated with allogeneic mature DC (104 cells per well) or by anti-CD3 (1 μg/ml) plus anti-CD28 mAb (2 μg/ml). Neutralizing anti-TGF-β mAb (10 μg/ml) was added to the cocultured cells as indicated. 3[H]Tdr was added after 3 (polyclonal stimulation) or 4 d (allogeneic MLR) of culture for the final 16 h.
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fig1: The suppressive activity of human CD25+ Treg cells is independent from membrane-bound TGF-β. CD4+ and CD4+CD25+ T cells were isolated from buffy coats of healthy volunteers by positive selection using paramagnetic beads. (A) Surface expression of TGF-β by freshly isolated CD4+ T cells and CD25+ Treg cells in comparison to the same T cell populations preactivated for 48 h with anti-CD3 (OKT3, 1 μg/ml) and anti-CD28 mAb (CD28.2, 2 μg/ml). The figure shows the expression of TGF-β (LAP-biotinylated) detected by streptavidin-Cy5 and CD4 (RPAT4-FITC). (B) CD4+ T cells (105 cells per well) or CD25+ Treg cells (105 cells per well), alone or in coculture (1:1), were stimulated with allogeneic mature DC (104 cells per well) or by anti-CD3 (1 μg/ml) plus anti-CD28 mAb (2 μg/ml). Neutralizing anti-TGF-β mAb (10 μg/ml) was added to the cocultured cells as indicated. 3[H]Tdr was added after 3 (polyclonal stimulation) or 4 d (allogeneic MLR) of culture for the final 16 h.

Mentions: CD25− conventional CD4+ T cells and CD25+ Treg cells were isolated from buffy coats of healthy volunteers as described previously (6). Freshly isolated, CD25+ Treg cells showed a fourfold increased expression of membrane-bound TGF-β as compared with conventional CD4+ Th cells (Fig. 1 A). However, polyclonal activation using anti-CD3 in combination with anti-CD28 antibodies resulted in an upregulated surface expression of TGF-β on conventional CD4+ T cells, whereas TGF-β on CD25+ Treg cells was downregulated. Both populations, CD25+ Treg cells and conventional CD4+ Th cells, either resting or activated, showed no significant production of biologically active soluble TGF-β (see Fig. 4 C).


Infectious tolerance: human CD25(+) regulatory T cells convey suppressor activity to conventional CD4(+) T helper cells.

Jonuleit H, Schmitt E, Kakirman H, Stassen M, Knop J, Enk AH - J. Exp. Med. (2002)

The suppressive activity of human CD25+ Treg cells is independent from membrane-bound TGF-β. CD4+ and CD4+CD25+ T cells were isolated from buffy coats of healthy volunteers by positive selection using paramagnetic beads. (A) Surface expression of TGF-β by freshly isolated CD4+ T cells and CD25+ Treg cells in comparison to the same T cell populations preactivated for 48 h with anti-CD3 (OKT3, 1 μg/ml) and anti-CD28 mAb (CD28.2, 2 μg/ml). The figure shows the expression of TGF-β (LAP-biotinylated) detected by streptavidin-Cy5 and CD4 (RPAT4-FITC). (B) CD4+ T cells (105 cells per well) or CD25+ Treg cells (105 cells per well), alone or in coculture (1:1), were stimulated with allogeneic mature DC (104 cells per well) or by anti-CD3 (1 μg/ml) plus anti-CD28 mAb (2 μg/ml). Neutralizing anti-TGF-β mAb (10 μg/ml) was added to the cocultured cells as indicated. 3[H]Tdr was added after 3 (polyclonal stimulation) or 4 d (allogeneic MLR) of culture for the final 16 h.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193929&req=5

fig1: The suppressive activity of human CD25+ Treg cells is independent from membrane-bound TGF-β. CD4+ and CD4+CD25+ T cells were isolated from buffy coats of healthy volunteers by positive selection using paramagnetic beads. (A) Surface expression of TGF-β by freshly isolated CD4+ T cells and CD25+ Treg cells in comparison to the same T cell populations preactivated for 48 h with anti-CD3 (OKT3, 1 μg/ml) and anti-CD28 mAb (CD28.2, 2 μg/ml). The figure shows the expression of TGF-β (LAP-biotinylated) detected by streptavidin-Cy5 and CD4 (RPAT4-FITC). (B) CD4+ T cells (105 cells per well) or CD25+ Treg cells (105 cells per well), alone or in coculture (1:1), were stimulated with allogeneic mature DC (104 cells per well) or by anti-CD3 (1 μg/ml) plus anti-CD28 mAb (2 μg/ml). Neutralizing anti-TGF-β mAb (10 μg/ml) was added to the cocultured cells as indicated. 3[H]Tdr was added after 3 (polyclonal stimulation) or 4 d (allogeneic MLR) of culture for the final 16 h.
Mentions: CD25− conventional CD4+ T cells and CD25+ Treg cells were isolated from buffy coats of healthy volunteers as described previously (6). Freshly isolated, CD25+ Treg cells showed a fourfold increased expression of membrane-bound TGF-β as compared with conventional CD4+ Th cells (Fig. 1 A). However, polyclonal activation using anti-CD3 in combination with anti-CD28 antibodies resulted in an upregulated surface expression of TGF-β on conventional CD4+ T cells, whereas TGF-β on CD25+ Treg cells was downregulated. Both populations, CD25+ Treg cells and conventional CD4+ Th cells, either resting or activated, showed no significant production of biologically active soluble TGF-β (see Fig. 4 C).

Bottom Line: Regulatory CD4(+)CD25(+) T cells (Treg) are mandatory for maintaining immunologic self-tolerance.Coactivation of CD25(+) Treg cells with Treg cell-depleted CD4(+) T cells results in anergized CD4(+) T cells that in turn inhibit the activation of conventional, freshly isolated CD4(+) T helper (Th) cells.This explains previously published conflicting data on the role of TGF-beta in CD25(+) Treg cell-induced immunosuppression.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Mainz, 55101 Mainz, Germany. jonuleit@hautklinik.klinik.uni-mainz.de

ABSTRACT
Regulatory CD4(+)CD25(+) T cells (Treg) are mandatory for maintaining immunologic self-tolerance. We demonstrate that the cell-cell contact-mediated suppression of conventional CD4(+) T cells by human CD25(+) Treg cells is fixation resistant, independent from membrane-bound TGF-beta but requires activation and protein synthesis of CD25(+) Treg cells. Coactivation of CD25(+) Treg cells with Treg cell-depleted CD4(+) T cells results in anergized CD4(+) T cells that in turn inhibit the activation of conventional, freshly isolated CD4(+) T helper (Th) cells. This infectious suppressive activity, transferred from CD25(+) Treg cells via cell contact, is cell contact-independent and partially mediated by soluble transforming growth factor (TGF)-beta. The induction of suppressive properties in conventional CD4(+) Th cells represents a mechanism underlying the phenomenon of infectious tolerance. This explains previously published conflicting data on the role of TGF-beta in CD25(+) Treg cell-induced immunosuppression.

Show MeSH
Related in: MedlinePlus