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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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Carboxy-PTIO inhibits the DPTA NONOate induction of IL-1β and IL-23.DCs were stimulated with Carboxy-PTIO (5μM) 30 min before treatment with DPTA NONOATE (0.6mM in RPMI or electrolyzed water (EW)). After 1h stimulation cells were maturated with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-1β, (B) IL-23 and (C) IL-12p70. Data are expressed as means ±SEM of 3 independent experiments. *P < 0.05.
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pone.0120134.g007: Carboxy-PTIO inhibits the DPTA NONOate induction of IL-1β and IL-23.DCs were stimulated with Carboxy-PTIO (5μM) 30 min before treatment with DPTA NONOATE (0.6mM in RPMI or electrolyzed water (EW)). After 1h stimulation cells were maturated with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-1β, (B) IL-23 and (C) IL-12p70. Data are expressed as means ±SEM of 3 independent experiments. *P < 0.05.

Mentions: In order to determine the specificity of NO, for the experiments, Carboxy-PTIO, was added to immature DC 30 min prior DPTA NONOate treatment. After 1 h incubation cells were matured with LPS. As shown in Fig. 7, the release of IL-23 induced by LPS or NO-LPS stimulation was markedly blocked by the addition of carboxy-PTIO (Fig. 7B). However, carboxy-PTIO markedly boosted the induction of IL-1β in both LPS and DPTA-Nonoate-LPS treatment (Fig. 7A). Carboxy-PTIO has been recognized as a specific NO scanvenger which directly oxidizes NO to form NO-2 and promote N2O3 formation [23]. Thus, the generation of these NO oxidative products may be involved in the IL-1β pathway activation, making difficult to determine the specificity of NO. In order to reduce active oxygen species generated during the co-stimulation with Carboxy-PTIO and DPTA NONOate, DPTA NONOate was dissolved in electrolyzed water (EW), which potently scavenges active oxygen species [22]. DPTA NONOate in EW release NO in a similar manner as in RPMI (S4 Fig). DPTA NONOate was added to DC pre treated or not with carboxy-PTIO and then matured with LPS. As a control 6 μL of EW was added to DC or LPS-DCs condition. DPTA NONOate in EW slightly decreases the release of IL-1β and IL-23 as compared to control conditions and maintains the inhibition of IL-12. Interestingly, in the presence of EW, carboxy-PTIO specifically inhibited the release of IL-1β and IL-23 without modifying the LPS response (Fig. 7A and B). In contrast, carboxy-PTIO in EW condition does not overcome the inhibitory effect of NO on the release of IL-12. Furthermore, carboxy-PTIO suppressed the LPS-release of IL-12 (Fig. 7C) suggesting that another unspecific effect of PTIO, which cannot be counteracted by EW, may act on the IL-12 pathway.


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)

Carboxy-PTIO inhibits the DPTA NONOate induction of IL-1β and IL-23.DCs were stimulated with Carboxy-PTIO (5μM) 30 min before treatment with DPTA NONOATE (0.6mM in RPMI or electrolyzed water (EW)). After 1h stimulation cells were maturated with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-1β, (B) IL-23 and (C) IL-12p70. Data are expressed as means ±SEM of 3 independent experiments. *P < 0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120134.g007: Carboxy-PTIO inhibits the DPTA NONOate induction of IL-1β and IL-23.DCs were stimulated with Carboxy-PTIO (5μM) 30 min before treatment with DPTA NONOATE (0.6mM in RPMI or electrolyzed water (EW)). After 1h stimulation cells were maturated with LPS (100ng/ml). The secretion of cytokines was analyzed using the Luminex system: (A) IL-1β, (B) IL-23 and (C) IL-12p70. Data are expressed as means ±SEM of 3 independent experiments. *P < 0.05.
Mentions: In order to determine the specificity of NO, for the experiments, Carboxy-PTIO, was added to immature DC 30 min prior DPTA NONOate treatment. After 1 h incubation cells were matured with LPS. As shown in Fig. 7, the release of IL-23 induced by LPS or NO-LPS stimulation was markedly blocked by the addition of carboxy-PTIO (Fig. 7B). However, carboxy-PTIO markedly boosted the induction of IL-1β in both LPS and DPTA-Nonoate-LPS treatment (Fig. 7A). Carboxy-PTIO has been recognized as a specific NO scanvenger which directly oxidizes NO to form NO-2 and promote N2O3 formation [23]. Thus, the generation of these NO oxidative products may be involved in the IL-1β pathway activation, making difficult to determine the specificity of NO. In order to reduce active oxygen species generated during the co-stimulation with Carboxy-PTIO and DPTA NONOate, DPTA NONOate was dissolved in electrolyzed water (EW), which potently scavenges active oxygen species [22]. DPTA NONOate in EW release NO in a similar manner as in RPMI (S4 Fig). DPTA NONOate was added to DC pre treated or not with carboxy-PTIO and then matured with LPS. As a control 6 μL of EW was added to DC or LPS-DCs condition. DPTA NONOate in EW slightly decreases the release of IL-1β and IL-23 as compared to control conditions and maintains the inhibition of IL-12. Interestingly, in the presence of EW, carboxy-PTIO specifically inhibited the release of IL-1β and IL-23 without modifying the LPS response (Fig. 7A and B). In contrast, carboxy-PTIO in EW condition does not overcome the inhibitory effect of NO on the release of IL-12. Furthermore, carboxy-PTIO suppressed the LPS-release of IL-12 (Fig. 7C) suggesting that another unspecific effect of PTIO, which cannot be counteracted by EW, may act on the IL-12 pathway.

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