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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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Th1 and Th2 cytokine production in peripheral T cells from nonpregnant and pregnant women. Peripheral blood mononuclear cells were isolated and stimulated with PMA/ionomycin. The percentage of CD4+/IFN-γ+, CD4+/TNF-α+, and CRTH2+/IL-4 positive cells were detected by flow cytometry in samples derived from nonpregnant (NP), 28-week pregnant, term no labour (TNL), and term labour (TL) samples (a), (c), and (e). A reduction in the percentage of peripheral CD4 positive cells secreting the Th1 cytokines IFN-γ and TNF-α was observed, whereas the percentage of cells secreting the Th2 cytokine, IL-4, remained consistent throughout gestation. Mean fluorescence intensity (MFI) was also assessed as a measure of total cytokine production (b), (d), and (f). PMA/ionomycin stimulation induced both Th1 and Th2 cytokine production in all sample groups compared to NL controls. No differences in gestation-dependent responses were detected. (g) a ratio of IFN-γ : IL-4 revealed a decrease in the Th1 : Th2 ratio during pregnancy, which was increased prior to the onset of labour. For statistical analysis ANOVA with Dunnett's multiple comparison test with NP as a control was used; **P < 0.01 and *P < 0.05.
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fig1: Th1 and Th2 cytokine production in peripheral T cells from nonpregnant and pregnant women. Peripheral blood mononuclear cells were isolated and stimulated with PMA/ionomycin. The percentage of CD4+/IFN-γ+, CD4+/TNF-α+, and CRTH2+/IL-4 positive cells were detected by flow cytometry in samples derived from nonpregnant (NP), 28-week pregnant, term no labour (TNL), and term labour (TL) samples (a), (c), and (e). A reduction in the percentage of peripheral CD4 positive cells secreting the Th1 cytokines IFN-γ and TNF-α was observed, whereas the percentage of cells secreting the Th2 cytokine, IL-4, remained consistent throughout gestation. Mean fluorescence intensity (MFI) was also assessed as a measure of total cytokine production (b), (d), and (f). PMA/ionomycin stimulation induced both Th1 and Th2 cytokine production in all sample groups compared to NL controls. No differences in gestation-dependent responses were detected. (g) a ratio of IFN-γ : IL-4 revealed a decrease in the Th1 : Th2 ratio during pregnancy, which was increased prior to the onset of labour. For statistical analysis ANOVA with Dunnett's multiple comparison test with NP as a control was used; **P < 0.01 and *P < 0.05.

Mentions: A change in the production of the Th1 and Th2 cytokines in pregnancy has previously been described. In this study, we employed flow cytometry to examine the effect of stimulation by the mitogen PMA/ionomycin on cytokine production at different gestational stages of pregnancy and during labour (see Figure 1). Th1 cytokine profiles of CD4 positive cells were assessed for intracellular IFN-γ and TNF-α (Figures 1(a)–1(d)) and compared to levels of nonpregnant controls. The percentage of peripheral T cells producing IFN-γ in response to stimulation reduced in pregnancy from 10.7% in nonpregnant women to 6.7% at 28 weeks, 5.1% at term (P < 0.05), and 5.6% at term in labour (P < 0.05). Similarly, the proportion of TNF-α producing cells was reduced, although not significantly, from 20.6% in nonpregnant women to 14.5% at 28 weeks, 15.8% at term and 13.3% at term in labour. Overall levels of Th1 cytokine production (expressed as mean fluorescence intensity), in the CD4+/IFN-γ+ or CD4+/TNF-α+ cells remained consistent throughout gestation and unchanged compared to nonpregnant controls. The Th2 cytokine, IL-4, was similarly assessed in CRTH2 positive cells (Figures 1(e) and 1(f)). While PMA/ionomycin stimulation did not increase the percentage of IL-4 expressing cells, the mean fluorescence intensity of IL-4 was significantly increased in samples collected from women at 28 weeks (39.3, P < 0.01) and at term (39.4, P < 0.05) compared to levels of nonpregnant controls (37.1). Levels of IL-4 in term labouring samples were consistent with nonlabouring samples (37.1). The ratio of the IFN-γ : IL-4 producing cells reduces during pregnancy, due to the suppression of the Th1 rather than the promotion of the Th2 cytokine production (Figure 1(g)).


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)

Th1 and Th2 cytokine production in peripheral T cells from nonpregnant and pregnant women. Peripheral blood mononuclear cells were isolated and stimulated with PMA/ionomycin. The percentage of CD4+/IFN-γ+, CD4+/TNF-α+, and CRTH2+/IL-4 positive cells were detected by flow cytometry in samples derived from nonpregnant (NP), 28-week pregnant, term no labour (TNL), and term labour (TL) samples (a), (c), and (e). A reduction in the percentage of peripheral CD4 positive cells secreting the Th1 cytokines IFN-γ and TNF-α was observed, whereas the percentage of cells secreting the Th2 cytokine, IL-4, remained consistent throughout gestation. Mean fluorescence intensity (MFI) was also assessed as a measure of total cytokine production (b), (d), and (f). PMA/ionomycin stimulation induced both Th1 and Th2 cytokine production in all sample groups compared to NL controls. No differences in gestation-dependent responses were detected. (g) a ratio of IFN-γ : IL-4 revealed a decrease in the Th1 : Th2 ratio during pregnancy, which was increased prior to the onset of labour. For statistical analysis ANOVA with Dunnett's multiple comparison test with NP as a control was used; **P < 0.01 and *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Th1 and Th2 cytokine production in peripheral T cells from nonpregnant and pregnant women. Peripheral blood mononuclear cells were isolated and stimulated with PMA/ionomycin. The percentage of CD4+/IFN-γ+, CD4+/TNF-α+, and CRTH2+/IL-4 positive cells were detected by flow cytometry in samples derived from nonpregnant (NP), 28-week pregnant, term no labour (TNL), and term labour (TL) samples (a), (c), and (e). A reduction in the percentage of peripheral CD4 positive cells secreting the Th1 cytokines IFN-γ and TNF-α was observed, whereas the percentage of cells secreting the Th2 cytokine, IL-4, remained consistent throughout gestation. Mean fluorescence intensity (MFI) was also assessed as a measure of total cytokine production (b), (d), and (f). PMA/ionomycin stimulation induced both Th1 and Th2 cytokine production in all sample groups compared to NL controls. No differences in gestation-dependent responses were detected. (g) a ratio of IFN-γ : IL-4 revealed a decrease in the Th1 : Th2 ratio during pregnancy, which was increased prior to the onset of labour. For statistical analysis ANOVA with Dunnett's multiple comparison test with NP as a control was used; **P < 0.01 and *P < 0.05.
Mentions: A change in the production of the Th1 and Th2 cytokines in pregnancy has previously been described. In this study, we employed flow cytometry to examine the effect of stimulation by the mitogen PMA/ionomycin on cytokine production at different gestational stages of pregnancy and during labour (see Figure 1). Th1 cytokine profiles of CD4 positive cells were assessed for intracellular IFN-γ and TNF-α (Figures 1(a)–1(d)) and compared to levels of nonpregnant controls. The percentage of peripheral T cells producing IFN-γ in response to stimulation reduced in pregnancy from 10.7% in nonpregnant women to 6.7% at 28 weeks, 5.1% at term (P < 0.05), and 5.6% at term in labour (P < 0.05). Similarly, the proportion of TNF-α producing cells was reduced, although not significantly, from 20.6% in nonpregnant women to 14.5% at 28 weeks, 15.8% at term and 13.3% at term in labour. Overall levels of Th1 cytokine production (expressed as mean fluorescence intensity), in the CD4+/IFN-γ+ or CD4+/TNF-α+ cells remained consistent throughout gestation and unchanged compared to nonpregnant controls. The Th2 cytokine, IL-4, was similarly assessed in CRTH2 positive cells (Figures 1(e) and 1(f)). While PMA/ionomycin stimulation did not increase the percentage of IL-4 expressing cells, the mean fluorescence intensity of IL-4 was significantly increased in samples collected from women at 28 weeks (39.3, P < 0.01) and at term (39.4, P < 0.05) compared to levels of nonpregnant controls (37.1). Levels of IL-4 in term labouring samples were consistent with nonlabouring samples (37.1). The ratio of the IFN-γ : IL-4 producing cells reduces during pregnancy, due to the suppression of the Th1 rather than the promotion of the Th2 cytokine production (Figure 1(g)).

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