Limits...
Cell contact-dependent immunosuppression by CD4(+)CD25(+) regulatory T cells is mediated by cell surface-bound transforming growth factor beta.

Nakamura K, Kitani A, Strober W - J. Exp. Med. (2001)

Bottom Line: In this study, we demonstrated that CD4(+)CD25(+) T cells do produce high levels of transforming growth factor (TGF)-beta1 and interleukin (IL)-10 compared with CD4(+)CD25(-) T cells when stimulated by plate-bound anti-CD3 and soluble anti-CD28 and/or IL-2, and secretion of TGF-beta1 (but not other cytokines), is further enhanced by costimulation via cytotoxic T lymphocyte-associated antigen (CTLA)-4.In addition, we found that CD4(+)CD25(+) T cells suppress B cell immunoglobulin production and that anti-TGF-beta again abolishes such suppression.Finally, we found that stimulated CD4(+)CD25(+) T cells but not CD4(+)CD25(-) T cells express high and persistent levels of TGF-beta1 on the cell surface.

View Article: PubMed Central - PubMed

Affiliation: Mucosal Immunity Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
CD4(+)CD25(+) T cells have been identified as a population of immunoregulatory T cells, which mediate suppression of CD4(+)CD25(-) T cells by cell-cell contact and not secretion of suppressor cytokines. In this study, we demonstrated that CD4(+)CD25(+) T cells do produce high levels of transforming growth factor (TGF)-beta1 and interleukin (IL)-10 compared with CD4(+)CD25(-) T cells when stimulated by plate-bound anti-CD3 and soluble anti-CD28 and/or IL-2, and secretion of TGF-beta1 (but not other cytokines), is further enhanced by costimulation via cytotoxic T lymphocyte-associated antigen (CTLA)-4. As in prior studies, we found that CD4(+)CD25(+) T cells suppress proliferation of CD4(+)CD25(-) T cells; however, we observed here that such suppression is abolished by the presence of anti-TGF-beta. In addition, we found that CD4(+)CD25(+) T cells suppress B cell immunoglobulin production and that anti-TGF-beta again abolishes such suppression. Finally, we found that stimulated CD4(+)CD25(+) T cells but not CD4(+)CD25(-) T cells express high and persistent levels of TGF-beta1 on the cell surface. This, plus the fact that we could find no evidence that a soluble factor mediates suppression, strongly suggests that CD4(+)CD25(+) T cells exert immunosuppression by a cell-cell interaction involving cell surface TGF-beta1.

Show MeSH

Related in: MedlinePlus

CD4+CD25+ T cells express LAP of TGF-β1 but not IL-10 or IL-4 on the cell surface. Purified CD4+CD25+ and CD4+CD25− T cells were stimulated with soluble anti-CD3 (10 μg/ml) and irradiated non–T cells in the presence of IL-2 (20 U/ml) for 24 h. Cells were incubated with Cy-Chrome™–conjugated anti-CD4 and either anti-LAP mAB (27232.11) (A), PE-conjugated anti–IL-10 (B), or PE-conjugated anti–IL-4 (C). Incubation with isotype-matched control IgG for each anti-cytokine was performed in parallel. In panel A, after incubation with anti-LAP, cells were washed, incubated with biotin-conjugated anti–mouse IgG1, washed, and incubated with PE-conjugated streptavidin. CD4+ cells were gated and expression of LAP (A), IL-10 (B), and IL-4 (C) on the cell surface was shown. Thick lines: anti-cytokine; thin lines: isotype-matched control IgG.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2195935&req=5

Figure 7: CD4+CD25+ T cells express LAP of TGF-β1 but not IL-10 or IL-4 on the cell surface. Purified CD4+CD25+ and CD4+CD25− T cells were stimulated with soluble anti-CD3 (10 μg/ml) and irradiated non–T cells in the presence of IL-2 (20 U/ml) for 24 h. Cells were incubated with Cy-Chrome™–conjugated anti-CD4 and either anti-LAP mAB (27232.11) (A), PE-conjugated anti–IL-10 (B), or PE-conjugated anti–IL-4 (C). Incubation with isotype-matched control IgG for each anti-cytokine was performed in parallel. In panel A, after incubation with anti-LAP, cells were washed, incubated with biotin-conjugated anti–mouse IgG1, washed, and incubated with PE-conjugated streptavidin. CD4+ cells were gated and expression of LAP (A), IL-10 (B), and IL-4 (C) on the cell surface was shown. Thick lines: anti-cytokine; thin lines: isotype-matched control IgG.

Mentions: Anti–TGF-β used in the above studies to detect cell surface TGF-β is a polyclonal (chicken) Ab raised against recombinant active (human) TGF-β1 and purified by TGF-β1 affinity chromatography; thus epitopes recognized by this Ab reside in active TGF-β1. To determine if latent TGF-β1 is in fact present on the cell surface, we also stained cells with Abs specific for LAP of TGF-β1. As shown in Fig. 7 A, we observed high levels of LAP expressed on the cell surface of stimulated CD4+CD25+ T cells using an anti-LAP mAb (clone 27232.11 [R&D Systems]) but only small amounts of LAP on the surface of stimulated CD4+CD25− T cells. Similar results were obtained with a polyclonal goat anti-LAP Ab (R&D Systems; data not shown). Finally, as shown in Fig. 7B and Fig. C, in contrast to the positive staining obtained with anti–TGF-β Ab or anti-LAP Abs, staining with anti–IL-10 or anti–IL-4 Abs for surface IL-10 or IL-4 was negative.


Cell contact-dependent immunosuppression by CD4(+)CD25(+) regulatory T cells is mediated by cell surface-bound transforming growth factor beta.

Nakamura K, Kitani A, Strober W - J. Exp. Med. (2001)

CD4+CD25+ T cells express LAP of TGF-β1 but not IL-10 or IL-4 on the cell surface. Purified CD4+CD25+ and CD4+CD25− T cells were stimulated with soluble anti-CD3 (10 μg/ml) and irradiated non–T cells in the presence of IL-2 (20 U/ml) for 24 h. Cells were incubated with Cy-Chrome™–conjugated anti-CD4 and either anti-LAP mAB (27232.11) (A), PE-conjugated anti–IL-10 (B), or PE-conjugated anti–IL-4 (C). Incubation with isotype-matched control IgG for each anti-cytokine was performed in parallel. In panel A, after incubation with anti-LAP, cells were washed, incubated with biotin-conjugated anti–mouse IgG1, washed, and incubated with PE-conjugated streptavidin. CD4+ cells were gated and expression of LAP (A), IL-10 (B), and IL-4 (C) on the cell surface was shown. Thick lines: anti-cytokine; thin lines: isotype-matched control IgG.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195935&req=5

Figure 7: CD4+CD25+ T cells express LAP of TGF-β1 but not IL-10 or IL-4 on the cell surface. Purified CD4+CD25+ and CD4+CD25− T cells were stimulated with soluble anti-CD3 (10 μg/ml) and irradiated non–T cells in the presence of IL-2 (20 U/ml) for 24 h. Cells were incubated with Cy-Chrome™–conjugated anti-CD4 and either anti-LAP mAB (27232.11) (A), PE-conjugated anti–IL-10 (B), or PE-conjugated anti–IL-4 (C). Incubation with isotype-matched control IgG for each anti-cytokine was performed in parallel. In panel A, after incubation with anti-LAP, cells were washed, incubated with biotin-conjugated anti–mouse IgG1, washed, and incubated with PE-conjugated streptavidin. CD4+ cells were gated and expression of LAP (A), IL-10 (B), and IL-4 (C) on the cell surface was shown. Thick lines: anti-cytokine; thin lines: isotype-matched control IgG.
Mentions: Anti–TGF-β used in the above studies to detect cell surface TGF-β is a polyclonal (chicken) Ab raised against recombinant active (human) TGF-β1 and purified by TGF-β1 affinity chromatography; thus epitopes recognized by this Ab reside in active TGF-β1. To determine if latent TGF-β1 is in fact present on the cell surface, we also stained cells with Abs specific for LAP of TGF-β1. As shown in Fig. 7 A, we observed high levels of LAP expressed on the cell surface of stimulated CD4+CD25+ T cells using an anti-LAP mAb (clone 27232.11 [R&D Systems]) but only small amounts of LAP on the surface of stimulated CD4+CD25− T cells. Similar results were obtained with a polyclonal goat anti-LAP Ab (R&D Systems; data not shown). Finally, as shown in Fig. 7B and Fig. C, in contrast to the positive staining obtained with anti–TGF-β Ab or anti-LAP Abs, staining with anti–IL-10 or anti–IL-4 Abs for surface IL-10 or IL-4 was negative.

Bottom Line: In this study, we demonstrated that CD4(+)CD25(+) T cells do produce high levels of transforming growth factor (TGF)-beta1 and interleukin (IL)-10 compared with CD4(+)CD25(-) T cells when stimulated by plate-bound anti-CD3 and soluble anti-CD28 and/or IL-2, and secretion of TGF-beta1 (but not other cytokines), is further enhanced by costimulation via cytotoxic T lymphocyte-associated antigen (CTLA)-4.In addition, we found that CD4(+)CD25(+) T cells suppress B cell immunoglobulin production and that anti-TGF-beta again abolishes such suppression.Finally, we found that stimulated CD4(+)CD25(+) T cells but not CD4(+)CD25(-) T cells express high and persistent levels of TGF-beta1 on the cell surface.

View Article: PubMed Central - PubMed

Affiliation: Mucosal Immunity Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
CD4(+)CD25(+) T cells have been identified as a population of immunoregulatory T cells, which mediate suppression of CD4(+)CD25(-) T cells by cell-cell contact and not secretion of suppressor cytokines. In this study, we demonstrated that CD4(+)CD25(+) T cells do produce high levels of transforming growth factor (TGF)-beta1 and interleukin (IL)-10 compared with CD4(+)CD25(-) T cells when stimulated by plate-bound anti-CD3 and soluble anti-CD28 and/or IL-2, and secretion of TGF-beta1 (but not other cytokines), is further enhanced by costimulation via cytotoxic T lymphocyte-associated antigen (CTLA)-4. As in prior studies, we found that CD4(+)CD25(+) T cells suppress proliferation of CD4(+)CD25(-) T cells; however, we observed here that such suppression is abolished by the presence of anti-TGF-beta. In addition, we found that CD4(+)CD25(+) T cells suppress B cell immunoglobulin production and that anti-TGF-beta again abolishes such suppression. Finally, we found that stimulated CD4(+)CD25(+) T cells but not CD4(+)CD25(-) T cells express high and persistent levels of TGF-beta1 on the cell surface. This, plus the fact that we could find no evidence that a soluble factor mediates suppression, strongly suggests that CD4(+)CD25(+) T cells exert immunosuppression by a cell-cell interaction involving cell surface TGF-beta1.

Show MeSH
Related in: MedlinePlus