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Exposure to inflammatory cytokines selectively limits GM-CSF production by induced T regulatory cells.

Reynolds BC, Turner DG, McPherson RC, Prendergast CT, Phelps RG, Turner NA, O'Connor RA, Anderton SM - Eur. J. Immunol. (2014)

Bottom Line: Understanding processes that can limit this potentially deleterious effect of Treg cells in a therapeutic setting is therefore important.We show that iTreg cells can produce significant amounts of three proinflammatory cytokines (IFN-γ, GM-CSF and TNF-α) upon secondary TCR stimulation.Furthermore, we show that IL-6 and IL-27 individually, or IL-2 and TGF-β in combination, can mediate the selective loss of GM-CSF production by iTreg cells.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Inflammation Research, University of Edinburgh, Edinburgh, UK.

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Related in: MedlinePlus

Production of IFN-γ, GM-CSF, and TNF-α is nonessential for iTreg-cell suppressive capacity. Varied numbers of iTreg cells were cocultured with a fixed number of naïve T responder cells as described in the Materials and methods. Proliferation was quantified by 3 H-thymidine incorporation after 96 h culture. Percent suppression was calculated, based on the means of triplicate cultures, following the addition, at the onset of co-culture, of (A) anti-TNF-α, (B) anti-GM-CSF, and (C) anti-IFN-γ. Filled triangles: anti-cytokine, open squares: isotype control antibodies. (D) The suppressive capacity of iTreg cells generated from Tg4.IFN-γ–/– mice were compared with that of iTreg cells generated from Tg4.CD90.1 (Tg4.WT) mice in suppression assays using naïve CD4+ Tg4 cells stimulated with MBP Ac1–9 and splenic APCs. Data are from one experiment representative of two experiments.
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fig03: Production of IFN-γ, GM-CSF, and TNF-α is nonessential for iTreg-cell suppressive capacity. Varied numbers of iTreg cells were cocultured with a fixed number of naïve T responder cells as described in the Materials and methods. Proliferation was quantified by 3 H-thymidine incorporation after 96 h culture. Percent suppression was calculated, based on the means of triplicate cultures, following the addition, at the onset of co-culture, of (A) anti-TNF-α, (B) anti-GM-CSF, and (C) anti-IFN-γ. Filled triangles: anti-cytokine, open squares: isotype control antibodies. (D) The suppressive capacity of iTreg cells generated from Tg4.IFN-γ–/– mice were compared with that of iTreg cells generated from Tg4.CD90.1 (Tg4.WT) mice in suppression assays using naïve CD4+ Tg4 cells stimulated with MBP Ac1–9 and splenic APCs. Data are from one experiment representative of two experiments.

Mentions: Thus far we had demonstrated that iTreg cells generated using a well characterized and widely used method would produce three proinflammatory cytokines upon secondary stimulation. We asked whether this would either diminish, or enhance, the strength of iTreg cell function using in vitro assays for suppression of naïve T cell activation. Although production of all three cytokines was again evident (data not shown), there was no apparent influence on the suppressive function of iTreg cells upon the proliferative response of naive T responder cells, stimulated by peptide-bearing APCs. Antibody neutralization of individual cytokines did not boost, or reduce, the observed suppression (Fig. 3A–C). Furthermore, IFN-γ-deficient iTreg did not have enhanced, or reduced, suppressive activity (Fig. 3D).


Exposure to inflammatory cytokines selectively limits GM-CSF production by induced T regulatory cells.

Reynolds BC, Turner DG, McPherson RC, Prendergast CT, Phelps RG, Turner NA, O'Connor RA, Anderton SM - Eur. J. Immunol. (2014)

Production of IFN-γ, GM-CSF, and TNF-α is nonessential for iTreg-cell suppressive capacity. Varied numbers of iTreg cells were cocultured with a fixed number of naïve T responder cells as described in the Materials and methods. Proliferation was quantified by 3 H-thymidine incorporation after 96 h culture. Percent suppression was calculated, based on the means of triplicate cultures, following the addition, at the onset of co-culture, of (A) anti-TNF-α, (B) anti-GM-CSF, and (C) anti-IFN-γ. Filled triangles: anti-cytokine, open squares: isotype control antibodies. (D) The suppressive capacity of iTreg cells generated from Tg4.IFN-γ–/– mice were compared with that of iTreg cells generated from Tg4.CD90.1 (Tg4.WT) mice in suppression assays using naïve CD4+ Tg4 cells stimulated with MBP Ac1–9 and splenic APCs. Data are from one experiment representative of two experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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fig03: Production of IFN-γ, GM-CSF, and TNF-α is nonessential for iTreg-cell suppressive capacity. Varied numbers of iTreg cells were cocultured with a fixed number of naïve T responder cells as described in the Materials and methods. Proliferation was quantified by 3 H-thymidine incorporation after 96 h culture. Percent suppression was calculated, based on the means of triplicate cultures, following the addition, at the onset of co-culture, of (A) anti-TNF-α, (B) anti-GM-CSF, and (C) anti-IFN-γ. Filled triangles: anti-cytokine, open squares: isotype control antibodies. (D) The suppressive capacity of iTreg cells generated from Tg4.IFN-γ–/– mice were compared with that of iTreg cells generated from Tg4.CD90.1 (Tg4.WT) mice in suppression assays using naïve CD4+ Tg4 cells stimulated with MBP Ac1–9 and splenic APCs. Data are from one experiment representative of two experiments.
Mentions: Thus far we had demonstrated that iTreg cells generated using a well characterized and widely used method would produce three proinflammatory cytokines upon secondary stimulation. We asked whether this would either diminish, or enhance, the strength of iTreg cell function using in vitro assays for suppression of naïve T cell activation. Although production of all three cytokines was again evident (data not shown), there was no apparent influence on the suppressive function of iTreg cells upon the proliferative response of naive T responder cells, stimulated by peptide-bearing APCs. Antibody neutralization of individual cytokines did not boost, or reduce, the observed suppression (Fig. 3A–C). Furthermore, IFN-γ-deficient iTreg did not have enhanced, or reduced, suppressive activity (Fig. 3D).

Bottom Line: Understanding processes that can limit this potentially deleterious effect of Treg cells in a therapeutic setting is therefore important.We show that iTreg cells can produce significant amounts of three proinflammatory cytokines (IFN-γ, GM-CSF and TNF-α) upon secondary TCR stimulation.Furthermore, we show that IL-6 and IL-27 individually, or IL-2 and TGF-β in combination, can mediate the selective loss of GM-CSF production by iTreg cells.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Inflammation Research, University of Edinburgh, Edinburgh, UK.

Show MeSH
Related in: MedlinePlus