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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. modulates the gene expression of IL-12, IL -23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) for 30 min before maturation with LPS (100ng/ml). Cells were analyzed at the different time-points indicated. (A) IL-12Bp40, (B) IL-12Ap35, (C) IL-23Ap19, (D) IL-1β and(E) IL-6 levels were assessed by TaqMan real-time reverse transcription—polymerase chain reaction (RT-PCR), using β-actin as endogenous control. Data express the relative gene abundance compared to control conditions, and are shown as mean ±SEM of 4 independent experiments, done in duplicates. *P < 0.05 or **P < 0.01.
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pone.0120134.g005: NO. modulates the gene expression of IL-12, IL -23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) for 30 min before maturation with LPS (100ng/ml). Cells were analyzed at the different time-points indicated. (A) IL-12Bp40, (B) IL-12Ap35, (C) IL-23Ap19, (D) IL-1β and(E) IL-6 levels were assessed by TaqMan real-time reverse transcription—polymerase chain reaction (RT-PCR), using β-actin as endogenous control. Data express the relative gene abundance compared to control conditions, and are shown as mean ±SEM of 4 independent experiments, done in duplicates. *P < 0.05 or **P < 0.01.

Mentions: To investigate the early events that modulate cytokine expression by NO. during the maturation of DCs, we measured the kinetics of the expression of mRNA encoding for IL-12Bp40, IL-12Ap35, IL-23Ap19, IL-1βand IL-IL-6 of DCs over 20 hours (Fig. 5). DPTA NONOate added 30 min before LPS stimulation has a strong inhibitory effect on the expression of IL-12p40 during the first 6 hours of LPS stimulation, but beyond 6 hours, the inhibition by DPTA NONOate seems to be lost (Fig. 5A). Interestingly, the expression of IL-12p35 at 6 hours was significantly increased (Fig. 5B), but the expression over 20 hours was reduced to levels similar measured in the presence of LPS alone. The cytokines involved in Th17 polarization were highly induced. IL-23p19 was increased after 6 hours and its expression is highly sustained for up to 20 hours when compared to LPS (Fig. 5C).


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. modulates the gene expression of IL-12, IL -23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) for 30 min before maturation with LPS (100ng/ml). Cells were analyzed at the different time-points indicated. (A) IL-12Bp40, (B) IL-12Ap35, (C) IL-23Ap19, (D) IL-1β and(E) IL-6 levels were assessed by TaqMan real-time reverse transcription—polymerase chain reaction (RT-PCR), using β-actin as endogenous control. Data express the relative gene abundance compared to control conditions, and are shown as mean ±SEM of 4 independent experiments, done in duplicates. *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.g005: NO. modulates the gene expression of IL-12, IL -23, IL-6 and IL-1β during maturation of DCs.DCs were stimulated with DPTA NONOate (0.6mM) for 30 min before maturation with LPS (100ng/ml). Cells were analyzed at the different time-points indicated. (A) IL-12Bp40, (B) IL-12Ap35, (C) IL-23Ap19, (D) IL-1β and(E) IL-6 levels were assessed by TaqMan real-time reverse transcription—polymerase chain reaction (RT-PCR), using β-actin as endogenous control. Data express the relative gene abundance compared to control conditions, and are shown as mean ±SEM of 4 independent experiments, done in duplicates. *P < 0.05 or **P < 0.01.
Mentions: To investigate the early events that modulate cytokine expression by NO. during the maturation of DCs, we measured the kinetics of the expression of mRNA encoding for IL-12Bp40, IL-12Ap35, IL-23Ap19, IL-1βand IL-IL-6 of DCs over 20 hours (Fig. 5). DPTA NONOate added 30 min before LPS stimulation has a strong inhibitory effect on the expression of IL-12p40 during the first 6 hours of LPS stimulation, but beyond 6 hours, the inhibition by DPTA NONOate seems to be lost (Fig. 5A). Interestingly, the expression of IL-12p35 at 6 hours was significantly increased (Fig. 5B), but the expression over 20 hours was reduced to levels similar measured in the presence of LPS alone. The cytokines involved in Th17 polarization were highly induced. IL-23p19 was increased after 6 hours and its expression is highly sustained for up to 20 hours when compared to LPS (Fig. 5C).

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