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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. inhibits the release of IL-12 at early time-points.DCs were stimulated with DPTA NONOATE (0.6mM) 10 min, 1h and 5h before maturation with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-10 (Due to a high variability of the IL-10 production the values were normalized to the levels of LPS %), (B) IL-12p70 and (C) TNFα. Data are expressed as means ±SEM of 11 independent experiments. *P < 0.05.
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pone.0120134.g002: NO. inhibits the release of IL-12 at early time-points.DCs were stimulated with DPTA NONOATE (0.6mM) 10 min, 1h and 5h before maturation with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-10 (Due to a high variability of the IL-10 production the values were normalized to the levels of LPS %), (B) IL-12p70 and (C) TNFα. Data are expressed as means ±SEM of 11 independent experiments. *P < 0.05.

Mentions: We next determined whether the duration of exposure to NO. before DC maturation can modulate the pattern of cytokine release by DCs. A concentration of 0.6mM was used for further experiments, since the toxicity of DPTA NONOate is low, with the ability to prevent apoptosis and spontaneous necrosis (S1 Fig.) and DPTA NONOate was also able to induce significant changes in cytokine release. DCs were stimulated with DPTA NONOate for 10 min, 1 or 5 hours before maturation with LPS to determine if an extended exposure to NO. was able to change the profile of cytokine release. When DCs were exposed to DPTA NONOate, the release of IL-12p70 and IL12-p40 was significantly reduced after only 10min incubation, but the inhibition was observed over the 5h period of incubation, compared to LPS-matured DCs (Fig. 2B and S2 Fig). DPTA NONOate did not modulate the release of IL-10 and TNF-α at any time-point (Fig. 2A and C). Thus, a short time pre-exposure to NO donor of 10 min before maturation with LPS was sufficient to convert DCs into an inhibitory role, modulating the inflammation induced by LPS, and decreasing the levels of IL-12. However, the 1- and 5-hour time periods were used for further experiments, since we considered it important that DCs should be exposed to NO. over long periods of time and because the strongest cytokine modulation was observed at these time-points.


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. inhibits the release of IL-12 at early time-points.DCs were stimulated with DPTA NONOATE (0.6mM) 10 min, 1h and 5h before maturation with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-10 (Due to a high variability of the IL-10 production the values were normalized to the levels of LPS %), (B) IL-12p70 and (C) TNFα. Data are expressed as means ±SEM of 11 independent experiments. *P < 0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120134.g002: NO. inhibits the release of IL-12 at early time-points.DCs were stimulated with DPTA NONOATE (0.6mM) 10 min, 1h and 5h before maturation with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-10 (Due to a high variability of the IL-10 production the values were normalized to the levels of LPS %), (B) IL-12p70 and (C) TNFα. Data are expressed as means ±SEM of 11 independent experiments. *P < 0.05.
Mentions: We next determined whether the duration of exposure to NO. before DC maturation can modulate the pattern of cytokine release by DCs. A concentration of 0.6mM was used for further experiments, since the toxicity of DPTA NONOate is low, with the ability to prevent apoptosis and spontaneous necrosis (S1 Fig.) and DPTA NONOate was also able to induce significant changes in cytokine release. DCs were stimulated with DPTA NONOate for 10 min, 1 or 5 hours before maturation with LPS to determine if an extended exposure to NO. was able to change the profile of cytokine release. When DCs were exposed to DPTA NONOate, the release of IL-12p70 and IL12-p40 was significantly reduced after only 10min incubation, but the inhibition was observed over the 5h period of incubation, compared to LPS-matured DCs (Fig. 2B and S2 Fig). DPTA NONOate did not modulate the release of IL-10 and TNF-α at any time-point (Fig. 2A and C). Thus, a short time pre-exposure to NO donor of 10 min before maturation with LPS was sufficient to convert DCs into an inhibitory role, modulating the inflammation induced by LPS, and decreasing the levels of IL-12. However, the 1- and 5-hour time periods were used for further experiments, since we considered it important that DCs should be exposed to NO. over long periods of time and because the strongest cytokine modulation was observed at these time-points.

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