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Activation of the D prostanoid 1 receptor suppresses asthma by modulation of lung dendritic cell function and induction of regulatory T cells.

Hammad H, Kool M, Soullié T, Narumiya S, Trottein F, Hoogsteden HC, Lambrecht BN - J. Exp. Med. (2007)

Bottom Line: Prostaglandins (PGs) can enhance or suppress inflammation by acting on different receptors expressed by hematopoietic and nonhematopoietic cells.These effects of DP1 agonist on DCs were mediated by cyclic AMP-dependent protein kinase A.Triggering DP1 on DCs is an important mechanism to induce regulatory T cells and to control the extent of airway inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary Medicine, Erasmus Medical Center, 3015 GE Rotterdam, Netherlands.

ABSTRACT
Prostaglandins (PGs) can enhance or suppress inflammation by acting on different receptors expressed by hematopoietic and nonhematopoietic cells. Prostaglandin D(2) binds to the D prostanoid (DP)1 and DP2 receptor and is seen as a critical mediator of asthma causing vasodilation, bronchoconstriction, and inflammatory cell influx. Here we show that inhalation of a selective DP1 agonist suppresses the cardinal features of asthma by targeting the function of lung dendritic cells (DCs). In mice treated with DP1 agonist or receiving DP1 agonist-treated DCs, there was an increase in Foxp3(+) CD4(+) regulatory T cells that suppressed inflammation in an interleukin 10-dependent way. These effects of DP1 agonist on DCs were mediated by cyclic AMP-dependent protein kinase A. We furthermore show that activation of DP1 by an endogenous ligand inhibits airway inflammation as chimeric mice with selective hematopoietic loss of DP1 had strongly enhanced airway inflammation and antigen-pulsed DCs lacking DP1 were better at inducing airway T helper 2 responses in the lung. Triggering DP1 on DCs is an important mechanism to induce regulatory T cells and to control the extent of airway inflammation. This pathway could be exploited to design novel treatments for asthma.

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Related in: MedlinePlus

BW245C treatment of DCs induces T reg cells. Vehicle/OVA-DCs, BW245C/OVA-DCs, and unpulsed DCs (Vehicle DCs) were cocultured for 4 d with naive OVA-specific CD4+ T cells. (A) Cell proliferation after a 16-h pulse. (B) Cytokines in the supernatant. (C) Foxp3 expression. (D) Regulatory T cell assay in vitro. Proliferation of OVA-specific T cells after a 2-d co-culture with syngeneic splenocytes and OVA peptide in the presence of T cells previously stimulated with vehicle/OVA-DCs (T-Vehicle/OVA-DCs) or BW245C/OVA-DCs (T-BW-DCs). (E) Regulatory T cell assay in vivo. On day 0, mice received an i.t. injection of vehicle/OVA-DCs, BW245C/OVA-DCs, or unpulsed DCs (Vehicle DCs). From days 10–13, mice were exposed to OVA aerosols. On day 14, CD4+ T cells were purified from MLNs and injected into recipient mice sensitized with OVA-alum 7 and 14 d earlier. 3 d after the CD4+ T cell transfer, mice were exposed to three OVA aerosols. BAL fluid was analyzed by flow cytometry. (F) The inhibitory effect of BW245C treatment of DCs is dependent on IL-10. On day 0, mice were injected i.t. with unpulsed DCs (Vehicle DCs), vehicle/OVA-DCs, or BW245C/OVA-DCs. Some mice were also injected i.p. with blocking anti–IL-10R antibodies 1 d before aerosol exposure. BAL fluid was analyzed by flow cytometry. Data are mean value ± SEM. *, P < 0.05. (G) Effects of BW245C on DCs depend on cAMP-dependent PKA. DCs were pulsed with OVA in the presence of vehicle, the PKA inhibitor Rp8-Br-cAMP, or the cell-permeable cAMP analogue 8-Br-cAMP, subsequently washed and put into culture with naive DO11.10 T cells. IL-10 production was measured 4 d later in supernatants. (H) Effects of BW245C in suppressing DC maturation (as indicated by CD40 up-regulation) depend on PKA and are mimicked by 8-Br-cAMP.
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fig6: BW245C treatment of DCs induces T reg cells. Vehicle/OVA-DCs, BW245C/OVA-DCs, and unpulsed DCs (Vehicle DCs) were cocultured for 4 d with naive OVA-specific CD4+ T cells. (A) Cell proliferation after a 16-h pulse. (B) Cytokines in the supernatant. (C) Foxp3 expression. (D) Regulatory T cell assay in vitro. Proliferation of OVA-specific T cells after a 2-d co-culture with syngeneic splenocytes and OVA peptide in the presence of T cells previously stimulated with vehicle/OVA-DCs (T-Vehicle/OVA-DCs) or BW245C/OVA-DCs (T-BW-DCs). (E) Regulatory T cell assay in vivo. On day 0, mice received an i.t. injection of vehicle/OVA-DCs, BW245C/OVA-DCs, or unpulsed DCs (Vehicle DCs). From days 10–13, mice were exposed to OVA aerosols. On day 14, CD4+ T cells were purified from MLNs and injected into recipient mice sensitized with OVA-alum 7 and 14 d earlier. 3 d after the CD4+ T cell transfer, mice were exposed to three OVA aerosols. BAL fluid was analyzed by flow cytometry. (F) The inhibitory effect of BW245C treatment of DCs is dependent on IL-10. On day 0, mice were injected i.t. with unpulsed DCs (Vehicle DCs), vehicle/OVA-DCs, or BW245C/OVA-DCs. Some mice were also injected i.p. with blocking anti–IL-10R antibodies 1 d before aerosol exposure. BAL fluid was analyzed by flow cytometry. Data are mean value ± SEM. *, P < 0.05. (G) Effects of BW245C on DCs depend on cAMP-dependent PKA. DCs were pulsed with OVA in the presence of vehicle, the PKA inhibitor Rp8-Br-cAMP, or the cell-permeable cAMP analogue 8-Br-cAMP, subsequently washed and put into culture with naive DO11.10 T cells. IL-10 production was measured 4 d later in supernatants. (H) Effects of BW245C in suppressing DC maturation (as indicated by CD40 up-regulation) depend on PKA and are mimicked by 8-Br-cAMP.

Mentions: The inhibitory effects of BW245C on induction of asthma suggested that DP1 ligation profoundly altered T cell polarization. To gain more information on this process, we next studied DC-driven T cell responses of OVA-specific (DO11.10) T cells in vitro. BW245C-treated OVA-DCs induced a lower proliferation and lower production of IL-5 in DO11.10 T cells compared with vehicle-treated OVA-DCs (Fig. 6, A and B), whereas the levels of the immunoregulatory cytokine IL-10 were strongly increased. The levels of IFN-γ induced by OVA-DC were unaffected by BW245C. Moreover, the percentage of CD4+25+Foxp3+ cells was increased when T cells were cocultured with BW245C/OVA-DCs compared with cells cocultured with vehicle/OVA-DCs (Fig. 6 C). As we found high levels of IL-10 induction and a higher number of Foxp3+ T cells with BW245C-treated OVA-DCs in vitro and in vivo (Fig. 1 D and Fig. 5 C) associated with a reduced proliferation of naive T cells, and we found increased numbers of Foxp3+CD25+ cells in vivo (Fig. 1 F and Fig. 5 C), we investigated whether BW245C-treated OVA-DCs induce the differentiation of T reg cells. As shown in Fig. 6 C, when OVA-DC–stimulated DO11.10 T cells were added to splenocytes and fresh naive DO11.10 T cells, OVA-specific T cell proliferation was stronger because of the presence of effector cells. However, BW245C/OVA-DC–stimulated T cells reduced the proliferation of fresh naive OVA-specific T cells in response to OVA-pulsed splenocytes.


Activation of the D prostanoid 1 receptor suppresses asthma by modulation of lung dendritic cell function and induction of regulatory T cells.

Hammad H, Kool M, Soullié T, Narumiya S, Trottein F, Hoogsteden HC, Lambrecht BN - J. Exp. Med. (2007)

BW245C treatment of DCs induces T reg cells. Vehicle/OVA-DCs, BW245C/OVA-DCs, and unpulsed DCs (Vehicle DCs) were cocultured for 4 d with naive OVA-specific CD4+ T cells. (A) Cell proliferation after a 16-h pulse. (B) Cytokines in the supernatant. (C) Foxp3 expression. (D) Regulatory T cell assay in vitro. Proliferation of OVA-specific T cells after a 2-d co-culture with syngeneic splenocytes and OVA peptide in the presence of T cells previously stimulated with vehicle/OVA-DCs (T-Vehicle/OVA-DCs) or BW245C/OVA-DCs (T-BW-DCs). (E) Regulatory T cell assay in vivo. On day 0, mice received an i.t. injection of vehicle/OVA-DCs, BW245C/OVA-DCs, or unpulsed DCs (Vehicle DCs). From days 10–13, mice were exposed to OVA aerosols. On day 14, CD4+ T cells were purified from MLNs and injected into recipient mice sensitized with OVA-alum 7 and 14 d earlier. 3 d after the CD4+ T cell transfer, mice were exposed to three OVA aerosols. BAL fluid was analyzed by flow cytometry. (F) The inhibitory effect of BW245C treatment of DCs is dependent on IL-10. On day 0, mice were injected i.t. with unpulsed DCs (Vehicle DCs), vehicle/OVA-DCs, or BW245C/OVA-DCs. Some mice were also injected i.p. with blocking anti–IL-10R antibodies 1 d before aerosol exposure. BAL fluid was analyzed by flow cytometry. Data are mean value ± SEM. *, P < 0.05. (G) Effects of BW245C on DCs depend on cAMP-dependent PKA. DCs were pulsed with OVA in the presence of vehicle, the PKA inhibitor Rp8-Br-cAMP, or the cell-permeable cAMP analogue 8-Br-cAMP, subsequently washed and put into culture with naive DO11.10 T cells. IL-10 production was measured 4 d later in supernatants. (H) Effects of BW245C in suppressing DC maturation (as indicated by CD40 up-regulation) depend on PKA and are mimicked by 8-Br-cAMP.
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Related In: Results  -  Collection

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fig6: BW245C treatment of DCs induces T reg cells. Vehicle/OVA-DCs, BW245C/OVA-DCs, and unpulsed DCs (Vehicle DCs) were cocultured for 4 d with naive OVA-specific CD4+ T cells. (A) Cell proliferation after a 16-h pulse. (B) Cytokines in the supernatant. (C) Foxp3 expression. (D) Regulatory T cell assay in vitro. Proliferation of OVA-specific T cells after a 2-d co-culture with syngeneic splenocytes and OVA peptide in the presence of T cells previously stimulated with vehicle/OVA-DCs (T-Vehicle/OVA-DCs) or BW245C/OVA-DCs (T-BW-DCs). (E) Regulatory T cell assay in vivo. On day 0, mice received an i.t. injection of vehicle/OVA-DCs, BW245C/OVA-DCs, or unpulsed DCs (Vehicle DCs). From days 10–13, mice were exposed to OVA aerosols. On day 14, CD4+ T cells were purified from MLNs and injected into recipient mice sensitized with OVA-alum 7 and 14 d earlier. 3 d after the CD4+ T cell transfer, mice were exposed to three OVA aerosols. BAL fluid was analyzed by flow cytometry. (F) The inhibitory effect of BW245C treatment of DCs is dependent on IL-10. On day 0, mice were injected i.t. with unpulsed DCs (Vehicle DCs), vehicle/OVA-DCs, or BW245C/OVA-DCs. Some mice were also injected i.p. with blocking anti–IL-10R antibodies 1 d before aerosol exposure. BAL fluid was analyzed by flow cytometry. Data are mean value ± SEM. *, P < 0.05. (G) Effects of BW245C on DCs depend on cAMP-dependent PKA. DCs were pulsed with OVA in the presence of vehicle, the PKA inhibitor Rp8-Br-cAMP, or the cell-permeable cAMP analogue 8-Br-cAMP, subsequently washed and put into culture with naive DO11.10 T cells. IL-10 production was measured 4 d later in supernatants. (H) Effects of BW245C in suppressing DC maturation (as indicated by CD40 up-regulation) depend on PKA and are mimicked by 8-Br-cAMP.
Mentions: The inhibitory effects of BW245C on induction of asthma suggested that DP1 ligation profoundly altered T cell polarization. To gain more information on this process, we next studied DC-driven T cell responses of OVA-specific (DO11.10) T cells in vitro. BW245C-treated OVA-DCs induced a lower proliferation and lower production of IL-5 in DO11.10 T cells compared with vehicle-treated OVA-DCs (Fig. 6, A and B), whereas the levels of the immunoregulatory cytokine IL-10 were strongly increased. The levels of IFN-γ induced by OVA-DC were unaffected by BW245C. Moreover, the percentage of CD4+25+Foxp3+ cells was increased when T cells were cocultured with BW245C/OVA-DCs compared with cells cocultured with vehicle/OVA-DCs (Fig. 6 C). As we found high levels of IL-10 induction and a higher number of Foxp3+ T cells with BW245C-treated OVA-DCs in vitro and in vivo (Fig. 1 D and Fig. 5 C) associated with a reduced proliferation of naive T cells, and we found increased numbers of Foxp3+CD25+ cells in vivo (Fig. 1 F and Fig. 5 C), we investigated whether BW245C-treated OVA-DCs induce the differentiation of T reg cells. As shown in Fig. 6 C, when OVA-DC–stimulated DO11.10 T cells were added to splenocytes and fresh naive DO11.10 T cells, OVA-specific T cell proliferation was stronger because of the presence of effector cells. However, BW245C/OVA-DC–stimulated T cells reduced the proliferation of fresh naive OVA-specific T cells in response to OVA-pulsed splenocytes.

Bottom Line: Prostaglandins (PGs) can enhance or suppress inflammation by acting on different receptors expressed by hematopoietic and nonhematopoietic cells.These effects of DP1 agonist on DCs were mediated by cyclic AMP-dependent protein kinase A.Triggering DP1 on DCs is an important mechanism to induce regulatory T cells and to control the extent of airway inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary Medicine, Erasmus Medical Center, 3015 GE Rotterdam, Netherlands.

ABSTRACT
Prostaglandins (PGs) can enhance or suppress inflammation by acting on different receptors expressed by hematopoietic and nonhematopoietic cells. Prostaglandin D(2) binds to the D prostanoid (DP)1 and DP2 receptor and is seen as a critical mediator of asthma causing vasodilation, bronchoconstriction, and inflammatory cell influx. Here we show that inhalation of a selective DP1 agonist suppresses the cardinal features of asthma by targeting the function of lung dendritic cells (DCs). In mice treated with DP1 agonist or receiving DP1 agonist-treated DCs, there was an increase in Foxp3(+) CD4(+) regulatory T cells that suppressed inflammation in an interleukin 10-dependent way. These effects of DP1 agonist on DCs were mediated by cyclic AMP-dependent protein kinase A. We furthermore show that activation of DP1 by an endogenous ligand inhibits airway inflammation as chimeric mice with selective hematopoietic loss of DP1 had strongly enhanced airway inflammation and antigen-pulsed DCs lacking DP1 were better at inducing airway T helper 2 responses in the lung. Triggering DP1 on DCs is an important mechanism to induce regulatory T cells and to control the extent of airway inflammation. This pathway could be exploited to design novel treatments for asthma.

Show MeSH
Related in: MedlinePlus