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Nitric oxide sustains IL-1β expression in human dendritic cells enhancing their capacity to induce IL-17-producing T-cells.

Obregon C, Graf L, Chung KF, Cesson V, Nicod LP - PLoS ONE (2015)

Bottom Line: NO changed the pattern of cytokine release by LPS-matured DCs, dependent on the concentration of NO, as well as on the timing of its addition to the cells during maturation.Indeed, DCs treated with NO efficiently induced the release of IL-17 by T-cells through IL-1β.Our work highlights the important role that NO may play in sustaining inflammation during an infection through the preferential differentiation of the Th17 lineage.

View Article: PubMed Central - PubMed

Affiliation: Pneumology Service, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland.

ABSTRACT
The role played by lung dendritic cells (DCs) which are influenced by external antigens and by their redox state in controlling inflammation is unclear. We studied the role played by nitric oxide (NO) in DC maturation and function. Human DCs were stimulated with a long-acting NO donor, DPTA NONOate, prior to exposure to lipopolysaccharide (LPS). Dose-and time-dependent experiments were performed with DCs with the aim of measuring the release and gene expression of inflammatory cytokines capable of modifying T-cell differentiation, towardsTh1, Th2 and Th17 cells. NO changed the pattern of cytokine release by LPS-matured DCs, dependent on the concentration of NO, as well as on the timing of its addition to the cells during maturation. Addition of NO before LPS-induced maturation strongly inhibited the release of IL-12, while increasing the expression and release of IL-23, IL-1β and IL-6, which are all involved in Th17 polarization. Indeed, DCs treated with NO efficiently induced the release of IL-17 by T-cells through IL-1β. Our work highlights the important role that NO may play in sustaining inflammation during an infection through the preferential differentiation of the Th17 lineage.

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NO. promotes the release of IL-23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) treated DCs 1h or 5h before maturation with LPS (100ng/ml). (A) IL-23 and (B) IL-6 were analyzed in supernatants. (C) IL-1β was analyzed in supernatants, but also in DCs pre-treated with the specific caspase-I inhibitor II Ac-YVAD-CMK (50 M) for 1 h before the DPTA NONOate /LPS stimulation. (D) Cells were harvested and intracellular pro-IL-1β (blotted with a specific pro-IL1β was analyzed by Western blot. Data are expressed as means ±SEM of 6 independent experiments. *P < 0.05 or **P < 0.01.
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pone.0120134.g004: NO. promotes the release of IL-23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) treated DCs 1h or 5h before maturation with LPS (100ng/ml). (A) IL-23 and (B) IL-6 were analyzed in supernatants. (C) IL-1β was analyzed in supernatants, but also in DCs pre-treated with the specific caspase-I inhibitor II Ac-YVAD-CMK (50 M) for 1 h before the DPTA NONOate /LPS stimulation. (D) Cells were harvested and intracellular pro-IL-1β (blotted with a specific pro-IL1β was analyzed by Western blot. Data are expressed as means ±SEM of 6 independent experiments. *P < 0.05 or **P < 0.01.

Mentions: To further characterize the role of NO. in the modulation of Th17 responses, we measured the production of the additional cytokines, IL-23, IL-6, TGF-β and IL-1β by DCs treated with NO. prior to addition of LPS. DPTA NONOate increased the release of IL-23 when added 1h before LPS (Fig. 4A). IL-6 was not modulated compared to LPS (Fig. 4B). The release of TGF-β was not increased compared to control conditions (unpublished data). Interestingly, IL-1βwas upregulated when compared to LPS alone, and this increase was sustained whether DPTA NONOate was used for 1 or 5 hours before LPS maturation (Fig. 4C). Different mechanisms of IL-1β release might be involved, since cell incubation in the presence of the specific caspase-1 inhibitor 1 hour before DPTA NONOate/LPS co-stimulation only inhibited the release when the cells were incubated with DPTA NONOate for 5 hours before LPS and not for 1 hour. Therefore, NO. may also be involved in the expression of IL-1β, since the pro-IL-1β is enhanced intracellularly (Fig. 4D). These results suggests that NO. is involved in the expression of IL-1β through at least 2 mechanisms. The expression of IL-1β is initially increased and, if NO. stimulation lasts longer, the cytokine release is increased through a caspase-1 dependent mechanism (Fig. 4C). Overall, NO. stimulated the release of IL-1β and IL-23, two cytokines that have been shown to be involved in the production of IL-17.


Nitric oxide sustains IL-1β expression in human dendritic cells enhancing their capacity to induce IL-17-producing T-cells.

Obregon C, Graf L, Chung KF, Cesson V, Nicod LP - PLoS ONE (2015)

NO. promotes the release of IL-23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) treated DCs 1h or 5h before maturation with LPS (100ng/ml). (A) IL-23 and (B) IL-6 were analyzed in supernatants. (C) IL-1β was analyzed in supernatants, but also in DCs pre-treated with the specific caspase-I inhibitor II Ac-YVAD-CMK (50 M) for 1 h before the DPTA NONOate /LPS stimulation. (D) Cells were harvested and intracellular pro-IL-1β (blotted with a specific pro-IL1β was analyzed by Western blot. Data are expressed as means ±SEM of 6 independent experiments. *P < 0.05 or **P < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390375&req=5

pone.0120134.g004: NO. promotes the release of IL-23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) treated DCs 1h or 5h before maturation with LPS (100ng/ml). (A) IL-23 and (B) IL-6 were analyzed in supernatants. (C) IL-1β was analyzed in supernatants, but also in DCs pre-treated with the specific caspase-I inhibitor II Ac-YVAD-CMK (50 M) for 1 h before the DPTA NONOate /LPS stimulation. (D) Cells were harvested and intracellular pro-IL-1β (blotted with a specific pro-IL1β was analyzed by Western blot. Data are expressed as means ±SEM of 6 independent experiments. *P < 0.05 or **P < 0.01.
Mentions: To further characterize the role of NO. in the modulation of Th17 responses, we measured the production of the additional cytokines, IL-23, IL-6, TGF-β and IL-1β by DCs treated with NO. prior to addition of LPS. DPTA NONOate increased the release of IL-23 when added 1h before LPS (Fig. 4A). IL-6 was not modulated compared to LPS (Fig. 4B). The release of TGF-β was not increased compared to control conditions (unpublished data). Interestingly, IL-1βwas upregulated when compared to LPS alone, and this increase was sustained whether DPTA NONOate was used for 1 or 5 hours before LPS maturation (Fig. 4C). Different mechanisms of IL-1β release might be involved, since cell incubation in the presence of the specific caspase-1 inhibitor 1 hour before DPTA NONOate/LPS co-stimulation only inhibited the release when the cells were incubated with DPTA NONOate for 5 hours before LPS and not for 1 hour. Therefore, NO. may also be involved in the expression of IL-1β, since the pro-IL-1β is enhanced intracellularly (Fig. 4D). These results suggests that NO. is involved in the expression of IL-1β through at least 2 mechanisms. The expression of IL-1β is initially increased and, if NO. stimulation lasts longer, the cytokine release is increased through a caspase-1 dependent mechanism (Fig. 4C). Overall, NO. stimulated the release of IL-1β and IL-23, two cytokines that have been shown to be involved in the production of IL-17.

Bottom Line: NO changed the pattern of cytokine release by LPS-matured DCs, dependent on the concentration of NO, as well as on the timing of its addition to the cells during maturation.Indeed, DCs treated with NO efficiently induced the release of IL-17 by T-cells through IL-1β.Our work highlights the important role that NO may play in sustaining inflammation during an infection through the preferential differentiation of the Th17 lineage.

View Article: PubMed Central - PubMed

Affiliation: Pneumology Service, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland.

ABSTRACT
The role played by lung dendritic cells (DCs) which are influenced by external antigens and by their redox state in controlling inflammation is unclear. We studied the role played by nitric oxide (NO) in DC maturation and function. Human DCs were stimulated with a long-acting NO donor, DPTA NONOate, prior to exposure to lipopolysaccharide (LPS). Dose-and time-dependent experiments were performed with DCs with the aim of measuring the release and gene expression of inflammatory cytokines capable of modifying T-cell differentiation, towardsTh1, Th2 and Th17 cells. NO changed the pattern of cytokine release by LPS-matured DCs, dependent on the concentration of NO, as well as on the timing of its addition to the cells during maturation. Addition of NO before LPS-induced maturation strongly inhibited the release of IL-12, while increasing the expression and release of IL-23, IL-1β and IL-6, which are all involved in Th17 polarization. Indeed, DCs treated with NO efficiently induced the release of IL-17 by T-cells through IL-1β. Our work highlights the important role that NO may play in sustaining inflammation during an infection through the preferential differentiation of the Th17 lineage.

Show MeSH
Related in: MedlinePlus