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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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Differential influence of IL-6, IL-12, and IL-27 upon cytokine production by Th0, Th1, and iTreg cells. Th0, Th1, and iTreg cells were generated from naive CD4+ Tg4.Foxp3LuciDTR-4 cells. Prior to restimulation, cells were FACS-sorted as GFP+ (for iTreg cells) or GFP− (for Th0 and Th1 cells). Cells were then restimulated for 48 h in triplicate with B10.PL splenic APCs and a dose response of MBP(Ac1–9) peptide with or without the addition of IL-6 (30 ng/mL), IL-12 (25 ng/mL), or IL-27 (50 ng/mL). Supernatants were analyzed by ELISA for production of the indicated cytokines. Data are shown as mean ± SEM of triplicates from one experiment representative of two experiments.
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fig06: Differential influence of IL-6, IL-12, and IL-27 upon cytokine production by Th0, Th1, and iTreg cells. Th0, Th1, and iTreg cells were generated from naive CD4+ Tg4.Foxp3LuciDTR-4 cells. Prior to restimulation, cells were FACS-sorted as GFP+ (for iTreg cells) or GFP− (for Th0 and Th1 cells). Cells were then restimulated for 48 h in triplicate with B10.PL splenic APCs and a dose response of MBP(Ac1–9) peptide with or without the addition of IL-6 (30 ng/mL), IL-12 (25 ng/mL), or IL-27 (50 ng/mL). Supernatants were analyzed by ELISA for production of the indicated cytokines. Data are shown as mean ± SEM of triplicates from one experiment representative of two experiments.

Mentions: We addressed whether the inhibitory effects of IL-6, IL-12, and IL-27 upon cytokine production were a unique feature of iTreg cells, or were common to other CD4+ T cell populations capable of producing IFN-γ, GM-CSF, and TNF-α (Fig. 6). For this comparison, iTreg, Th0, and Th1 cells were generated from naïve Tg4.Foxp3LuciDTR-4 cells and restimulated with splenic APCs and a dose response of MBP peptide. In the absence of exogenous cytokines, iTreg cells produced lower levels of GM-CSF and TNF-α than their Th0 and Th1 counterparts. Levels of IFN-γ were similar between iTreg cells and Th0 cells and, as would be expected, these were lower than those detected from Th1 cells (Fig. 6A–C). Addition of either IL-6 or IL-27 inhibited iTreg-cell production of GM-CSF (Fig. 6F), mirroring their effects seen with APC-free TCR stimulation (in Fig. 5). However, inhibition of GM-CSF was not seen with IL-12. GM-CSF production by iTreg cells and Th0 cells was affected in a similar way by exogenous cytokines (Fig. 6D and F), whereas no clear inhibition of Th1-cell production of GM-CSF was evident (Fig. 6E). The only cytokine to clearly elevate IFN-γ production by iTreg cells was IL-12; IL-27 could not achieve this (Fig. 6C). We conclude that, while there is some overlap in how IL-6, IL-12, and IL-27 influence cytokine production by Th0, Th1, and iTreg cells, each cell-type shows a unique response profile to these cytokines.


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)

Differential influence of IL-6, IL-12, and IL-27 upon cytokine production by Th0, Th1, and iTreg cells. Th0, Th1, and iTreg cells were generated from naive CD4+ Tg4.Foxp3LuciDTR-4 cells. Prior to restimulation, cells were FACS-sorted as GFP+ (for iTreg cells) or GFP− (for Th0 and Th1 cells). Cells were then restimulated for 48 h in triplicate with B10.PL splenic APCs and a dose response of MBP(Ac1–9) peptide with or without the addition of IL-6 (30 ng/mL), IL-12 (25 ng/mL), or IL-27 (50 ng/mL). Supernatants were analyzed by ELISA for production of the indicated cytokines. Data are shown as mean ± SEM of triplicates from one experiment representative of two experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4257504&req=5

fig06: Differential influence of IL-6, IL-12, and IL-27 upon cytokine production by Th0, Th1, and iTreg cells. Th0, Th1, and iTreg cells were generated from naive CD4+ Tg4.Foxp3LuciDTR-4 cells. Prior to restimulation, cells were FACS-sorted as GFP+ (for iTreg cells) or GFP− (for Th0 and Th1 cells). Cells were then restimulated for 48 h in triplicate with B10.PL splenic APCs and a dose response of MBP(Ac1–9) peptide with or without the addition of IL-6 (30 ng/mL), IL-12 (25 ng/mL), or IL-27 (50 ng/mL). Supernatants were analyzed by ELISA for production of the indicated cytokines. Data are shown as mean ± SEM of triplicates from one experiment representative of two experiments.
Mentions: We addressed whether the inhibitory effects of IL-6, IL-12, and IL-27 upon cytokine production were a unique feature of iTreg cells, or were common to other CD4+ T cell populations capable of producing IFN-γ, GM-CSF, and TNF-α (Fig. 6). For this comparison, iTreg, Th0, and Th1 cells were generated from naïve Tg4.Foxp3LuciDTR-4 cells and restimulated with splenic APCs and a dose response of MBP peptide. In the absence of exogenous cytokines, iTreg cells produced lower levels of GM-CSF and TNF-α than their Th0 and Th1 counterparts. Levels of IFN-γ were similar between iTreg cells and Th0 cells and, as would be expected, these were lower than those detected from Th1 cells (Fig. 6A–C). Addition of either IL-6 or IL-27 inhibited iTreg-cell production of GM-CSF (Fig. 6F), mirroring their effects seen with APC-free TCR stimulation (in Fig. 5). However, inhibition of GM-CSF was not seen with IL-12. GM-CSF production by iTreg cells and Th0 cells was affected in a similar way by exogenous cytokines (Fig. 6D and F), whereas no clear inhibition of Th1-cell production of GM-CSF was evident (Fig. 6E). The only cytokine to clearly elevate IFN-γ production by iTreg cells was IL-12; IL-27 could not achieve this (Fig. 6C). We conclude that, while there is some overlap in how IL-6, IL-12, and IL-27 influence cytokine production by Th0, Th1, and iTreg cells, each cell-type shows a unique response profile to these cytokines.

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