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Changes in the Th1:Th2 cytokine bias in pregnancy and the effects of the anti-inflammatory cyclopentenone prostaglandin 15-deoxy-Δ(12,14)-prostaglandin J2.

Sykes L, MacIntyre DA, Yap XJ, Ponnampalam S, Teoh TG, Bennett PR - Mediators Inflamm. (2012)

Bottom Line: The anti-inflammatory prostaglandin 15-deoxy-Δ(12,14)-Prostaglandin J(2) (15dPGJ(2)) inhibits nuclear factor Kappa B (NF-κB) in amniocytes and myocytes in vitro and is a ligand for the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) receptor.The percentage of CD4(+) cells producing interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in response to PMA/ionomycin was significantly reduced in pregnancy. 15dPGJ(2) reduced IFN-γ and TNF-α production in stimulated T helper cells, but did not alter IL-4 production in CRTH2(+ve) cells. 15dPGJ(2) also reduced phospho-p65 in stimulated PBMCs.In summary, 15dPGJ(2) suppresses the Th1 response of PBMCs during pregnancy and active labour whilst maintaining the Th2 response suggesting a therapeutic benefit in reducing neonatal morbidity in inflammation-induced PTL.

View Article: PubMed Central - PubMed

Affiliation: Parturition Research Group, Department of Surgery and Cancer, Institute of Reproduction and Developmental Biology, Imperial College London, W120NN London, UK.

ABSTRACT
Pregnancy is a complex immunological state in which a bias towards T helper 2 (Th2) protects the fetus. Evidence suggests that proinflammatory cytokines increase the risk of poor neonatal outcome, independently of the direct effect of preterm labour. The anti-inflammatory prostaglandin 15-deoxy-Δ(12,14)-Prostaglandin J(2) (15dPGJ(2)) inhibits nuclear factor Kappa B (NF-κB) in amniocytes and myocytes in vitro and is a ligand for the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) receptor. Here we examine the Th1:Th2 cytokine bias in pregnancy and whether 15dPGJ(2) could be used to inhibit the production of the proinflammatory cytokines through inhibition of NF-κB while simultaneously promoting Th2 interleukin 4 (IL-4) synthesis via CRTH2 in T helper cells. Peripheral blood mononuclear cells (PBMCs) from women at 28 weeks, term pre-labour, term labour as well as non-pregnant female controls were cultured with 15dPGJ(2) or vehicle control and stimulated with phorbol myristyl acetate (PMA)/ionomycin. The percentage of CD4(+) cells producing interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in response to PMA/ionomycin was significantly reduced in pregnancy. 15dPGJ(2) reduced IFN-γ and TNF-α production in stimulated T helper cells, but did not alter IL-4 production in CRTH2(+ve) cells. 15dPGJ(2) also reduced phospho-p65 in stimulated PBMCs. In summary, 15dPGJ(2) suppresses the Th1 response of PBMCs during pregnancy and active labour whilst maintaining the Th2 response suggesting a therapeutic benefit in reducing neonatal morbidity in inflammation-induced PTL.

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The effect of 15dPGJ2 on NF-κB p65 phosphorylation in peripheral blood mononuclear cells. PBMCs were treated as previously described before being extracted and levels of phosphorylated p65 (p-p65) examined using immunoblotting. Representative immunoblots are shown for each gestational time point (a). Immunoblots were reprobed for β-actin as an internal loading control. Densitometric analysis of the immunoblots was conducted revealing a significant increase in p-p65 levels upon stimulation with PMA/ionomycin in all samples (b). A significant decrease in PMA/ionomycin stimulated p-p65 was observed following 15dPGJ2 treatment in 28-week samples. The capacity of 15dPGJ2 to inhibit PMA/ionomycin-induced p-p65 was lost in those samples collected at term. NS: nonstimulated, S: PMA/ionomycin stimulated, and J2: 15dPGJ2 pretreatment. Effect of treatment was examined for statistical significance at each gestational timepoint using ANOVA with Bonferroni's multiple comparison test; ***P < 0.001, **P < 0.01 and *P < 0.05.
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fig4: The effect of 15dPGJ2 on NF-κB p65 phosphorylation in peripheral blood mononuclear cells. PBMCs were treated as previously described before being extracted and levels of phosphorylated p65 (p-p65) examined using immunoblotting. Representative immunoblots are shown for each gestational time point (a). Immunoblots were reprobed for β-actin as an internal loading control. Densitometric analysis of the immunoblots was conducted revealing a significant increase in p-p65 levels upon stimulation with PMA/ionomycin in all samples (b). A significant decrease in PMA/ionomycin stimulated p-p65 was observed following 15dPGJ2 treatment in 28-week samples. The capacity of 15dPGJ2 to inhibit PMA/ionomycin-induced p-p65 was lost in those samples collected at term. NS: nonstimulated, S: PMA/ionomycin stimulated, and J2: 15dPGJ2 pretreatment. Effect of treatment was examined for statistical significance at each gestational timepoint using ANOVA with Bonferroni's multiple comparison test; ***P < 0.001, **P < 0.01 and *P < 0.05.

Mentions: Peripheral blood mononuclear cells were incubated with vehicle control or 32 μM 15dPGJ2 and stimulated with PMA/ionomycin for 10 min before being extracted and assessed for levels of phosphorylated p65, the transcriptionally active subunit of NF-κB. PMA/ionomycin treatment stimulated NF-κB phosphorylation in all samples and at all gestational time points (Figure 4). Pretreatment of cells with 15dPGJ2 significantly reduced levels of p-p65 in samples collected from women at 28-week gestation. A clear trend of decreased p-p65 following 15dPGJ2 was observed in the remaining samples yet they did not return to basal levels of p-p65.


Changes in the Th1:Th2 cytokine bias in pregnancy and the effects of the anti-inflammatory cyclopentenone prostaglandin 15-deoxy-Δ(12,14)-prostaglandin J2.

Sykes L, MacIntyre DA, Yap XJ, Ponnampalam S, Teoh TG, Bennett PR - Mediators Inflamm. (2012)

The effect of 15dPGJ2 on NF-κB p65 phosphorylation in peripheral blood mononuclear cells. PBMCs were treated as previously described before being extracted and levels of phosphorylated p65 (p-p65) examined using immunoblotting. Representative immunoblots are shown for each gestational time point (a). Immunoblots were reprobed for β-actin as an internal loading control. Densitometric analysis of the immunoblots was conducted revealing a significant increase in p-p65 levels upon stimulation with PMA/ionomycin in all samples (b). A significant decrease in PMA/ionomycin stimulated p-p65 was observed following 15dPGJ2 treatment in 28-week samples. The capacity of 15dPGJ2 to inhibit PMA/ionomycin-induced p-p65 was lost in those samples collected at term. NS: nonstimulated, S: PMA/ionomycin stimulated, and J2: 15dPGJ2 pretreatment. Effect of treatment was examined for statistical significance at each gestational timepoint using ANOVA with Bonferroni's multiple comparison test; ***P < 0.001, **P < 0.01 and *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368617&req=5

fig4: The effect of 15dPGJ2 on NF-κB p65 phosphorylation in peripheral blood mononuclear cells. PBMCs were treated as previously described before being extracted and levels of phosphorylated p65 (p-p65) examined using immunoblotting. Representative immunoblots are shown for each gestational time point (a). Immunoblots were reprobed for β-actin as an internal loading control. Densitometric analysis of the immunoblots was conducted revealing a significant increase in p-p65 levels upon stimulation with PMA/ionomycin in all samples (b). A significant decrease in PMA/ionomycin stimulated p-p65 was observed following 15dPGJ2 treatment in 28-week samples. The capacity of 15dPGJ2 to inhibit PMA/ionomycin-induced p-p65 was lost in those samples collected at term. NS: nonstimulated, S: PMA/ionomycin stimulated, and J2: 15dPGJ2 pretreatment. Effect of treatment was examined for statistical significance at each gestational timepoint using ANOVA with Bonferroni's multiple comparison test; ***P < 0.001, **P < 0.01 and *P < 0.05.
Mentions: Peripheral blood mononuclear cells were incubated with vehicle control or 32 μM 15dPGJ2 and stimulated with PMA/ionomycin for 10 min before being extracted and assessed for levels of phosphorylated p65, the transcriptionally active subunit of NF-κB. PMA/ionomycin treatment stimulated NF-κB phosphorylation in all samples and at all gestational time points (Figure 4). Pretreatment of cells with 15dPGJ2 significantly reduced levels of p-p65 in samples collected from women at 28-week gestation. A clear trend of decreased p-p65 following 15dPGJ2 was observed in the remaining samples yet they did not return to basal levels of p-p65.

Bottom Line: The anti-inflammatory prostaglandin 15-deoxy-Δ(12,14)-Prostaglandin J(2) (15dPGJ(2)) inhibits nuclear factor Kappa B (NF-κB) in amniocytes and myocytes in vitro and is a ligand for the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) receptor.The percentage of CD4(+) cells producing interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in response to PMA/ionomycin was significantly reduced in pregnancy. 15dPGJ(2) reduced IFN-γ and TNF-α production in stimulated T helper cells, but did not alter IL-4 production in CRTH2(+ve) cells. 15dPGJ(2) also reduced phospho-p65 in stimulated PBMCs.In summary, 15dPGJ(2) suppresses the Th1 response of PBMCs during pregnancy and active labour whilst maintaining the Th2 response suggesting a therapeutic benefit in reducing neonatal morbidity in inflammation-induced PTL.

View Article: PubMed Central - PubMed

Affiliation: Parturition Research Group, Department of Surgery and Cancer, Institute of Reproduction and Developmental Biology, Imperial College London, W120NN London, UK.

ABSTRACT
Pregnancy is a complex immunological state in which a bias towards T helper 2 (Th2) protects the fetus. Evidence suggests that proinflammatory cytokines increase the risk of poor neonatal outcome, independently of the direct effect of preterm labour. The anti-inflammatory prostaglandin 15-deoxy-Δ(12,14)-Prostaglandin J(2) (15dPGJ(2)) inhibits nuclear factor Kappa B (NF-κB) in amniocytes and myocytes in vitro and is a ligand for the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) receptor. Here we examine the Th1:Th2 cytokine bias in pregnancy and whether 15dPGJ(2) could be used to inhibit the production of the proinflammatory cytokines through inhibition of NF-κB while simultaneously promoting Th2 interleukin 4 (IL-4) synthesis via CRTH2 in T helper cells. Peripheral blood mononuclear cells (PBMCs) from women at 28 weeks, term pre-labour, term labour as well as non-pregnant female controls were cultured with 15dPGJ(2) or vehicle control and stimulated with phorbol myristyl acetate (PMA)/ionomycin. The percentage of CD4(+) cells producing interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in response to PMA/ionomycin was significantly reduced in pregnancy. 15dPGJ(2) reduced IFN-γ and TNF-α production in stimulated T helper cells, but did not alter IL-4 production in CRTH2(+ve) cells. 15dPGJ(2) also reduced phospho-p65 in stimulated PBMCs. In summary, 15dPGJ(2) suppresses the Th1 response of PBMCs during pregnancy and active labour whilst maintaining the Th2 response suggesting a therapeutic benefit in reducing neonatal morbidity in inflammation-induced PTL.

Show MeSH
Related in: MedlinePlus