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Prostaglandin E2 and SOCS1 have a role in intestinal immune tolerance.

Chinen T, Komai K, Muto G, Morita R, Inoue N, Yoshida H, Sekiya T, Yoshida R, Nakamura K, Takayanagi R, Yoshimura A - Nat Commun (2011)

Bottom Line: Additionally, we find an essential role for prostaglandin E2 (PGE2) in the maintenance of tolerance within the intestine in the absence of Tregs.Socs1(-/-) dendritic cells are resistant to PGE2-mediated immunosuppression because of dysregulated cytokine signalling.Thus, we propose that SOCS1 and PGE2, potentially interacting together, act as an alternative intestinal tolerance mechanism distinct from IL-10 and Tregs.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

ABSTRACT
Interleukin 10 (IL-10) and regulatory T cells (Tregs) maintain tolerance to intestinal microorganisms. However, Il10(-/-)Rag2(-/-) mice, which lack IL-10 and Tregs, remain healthy, suggesting the existence of other mechanisms of tolerance. Here, we identify suppressor of cytokine signalling 1 (SOCS1) as an essential mediator of immune tolerance in the intestine. Socs1(-/-)Rag2(-/-) mice develop severe colitis, which can be prevented by the reduction of microbiota and the transfer of IL-10-sufficient Tregs. Additionally, we find an essential role for prostaglandin E2 (PGE2) in the maintenance of tolerance within the intestine in the absence of Tregs. Socs1(-/-) dendritic cells are resistant to PGE2-mediated immunosuppression because of dysregulated cytokine signalling. Thus, we propose that SOCS1 and PGE2, potentially interacting together, act as an alternative intestinal tolerance mechanism distinct from IL-10 and Tregs.

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IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1−/− BMDCs.(a) Socs1+/+ (5×105 cells per ml; black bars) and Socs1−/− BMDCs (5×105 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml−1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. (b) Socs1+/+ (5×105 cells per ml; left half of each panel) and Socs1−/− BMDCs (5×105 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml−1) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. (c) Stat1+/+ (5×105 cells per ml; white bars) and Stat1−/− (5×105 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml−1) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. (d) Socs1+/+ and Socs1−/− BMDCs (5×105 cells per ml) were co-cultured with 2.5×105 cells per ml of CD4+CD25high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with 10 ng ml−1 of LPS+IFNγ (10 ng ml−1, each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml−1) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two (b–d) to three (a) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. **P<0.01.
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f5: IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1−/− BMDCs.(a) Socs1+/+ (5×105 cells per ml; black bars) and Socs1−/− BMDCs (5×105 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml−1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. (b) Socs1+/+ (5×105 cells per ml; left half of each panel) and Socs1−/− BMDCs (5×105 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml−1) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. (c) Stat1+/+ (5×105 cells per ml; white bars) and Stat1−/− (5×105 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml−1) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. (d) Socs1+/+ and Socs1−/− BMDCs (5×105 cells per ml) were co-cultured with 2.5×105 cells per ml of CD4+CD25high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with 10 ng ml−1 of LPS+IFNγ (10 ng ml−1, each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml−1) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two (b–d) to three (a) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. **P<0.01.

Mentions: We attempted to ascertain how the PGE2-mediated immunosuppression was impaired in Socs1−/− BMDCs. As mentioned in Figure 3h, the suppressive effect of PGE2 on LPS+IFNγ-induced IL-12p70 and TNFα production from BMDCs was counteracted by SOCS1 deficiency, and this antagonism was barely seen in the absence of IFNγ, suggesting that dysregulated IFNγ signal is the cause of the antagonism. When PGE2 binds to a receptor, especially PTGER4, adenylate cyclase is activated and the level of cyclic AMP (cAMP) is elevated. cAMP then suppresses the production of inflammatory cytokines by several mechanisms1819. As shown in Figure 5a, Socs1−/− BMDCs were also resistant to cAMP-mediated suppression of cytokine production in the presence of LPS+IFNγ. IFNγ dose-dependently counteracted the suppressive effects of cAMP in Socs1−/− BMDCs (Fig. 5b). It is notable that IFNγ partially antagonized the suppressive effect of PGE2 and cAMP in Socs1+/+ BMDCs (Fig. 5b), although this antagonism was not observed in Stat1−/− BMDCs (Fig. 5c). These data suggest that the antagonistic effect of IFNγ/STAT1 pathway against PGE2 and cAMP was amplified in Socs1−/− BMDCs.


Prostaglandin E2 and SOCS1 have a role in intestinal immune tolerance.

Chinen T, Komai K, Muto G, Morita R, Inoue N, Yoshida H, Sekiya T, Yoshida R, Nakamura K, Takayanagi R, Yoshimura A - Nat Commun (2011)

IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1−/− BMDCs.(a) Socs1+/+ (5×105 cells per ml; black bars) and Socs1−/− BMDCs (5×105 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml−1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. (b) Socs1+/+ (5×105 cells per ml; left half of each panel) and Socs1−/− BMDCs (5×105 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml−1) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. (c) Stat1+/+ (5×105 cells per ml; white bars) and Stat1−/− (5×105 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml−1) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. (d) Socs1+/+ and Socs1−/− BMDCs (5×105 cells per ml) were co-cultured with 2.5×105 cells per ml of CD4+CD25high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with 10 ng ml−1 of LPS+IFNγ (10 ng ml−1, each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml−1) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two (b–d) to three (a) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. **P<0.01.
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f5: IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1−/− BMDCs.(a) Socs1+/+ (5×105 cells per ml; black bars) and Socs1−/− BMDCs (5×105 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml−1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. (b) Socs1+/+ (5×105 cells per ml; left half of each panel) and Socs1−/− BMDCs (5×105 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml−1) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. (c) Stat1+/+ (5×105 cells per ml; white bars) and Stat1−/− (5×105 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml−1) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. (d) Socs1+/+ and Socs1−/− BMDCs (5×105 cells per ml) were co-cultured with 2.5×105 cells per ml of CD4+CD25high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with 10 ng ml−1 of LPS+IFNγ (10 ng ml−1, each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml−1) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two (b–d) to three (a) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. **P<0.01.
Mentions: We attempted to ascertain how the PGE2-mediated immunosuppression was impaired in Socs1−/− BMDCs. As mentioned in Figure 3h, the suppressive effect of PGE2 on LPS+IFNγ-induced IL-12p70 and TNFα production from BMDCs was counteracted by SOCS1 deficiency, and this antagonism was barely seen in the absence of IFNγ, suggesting that dysregulated IFNγ signal is the cause of the antagonism. When PGE2 binds to a receptor, especially PTGER4, adenylate cyclase is activated and the level of cyclic AMP (cAMP) is elevated. cAMP then suppresses the production of inflammatory cytokines by several mechanisms1819. As shown in Figure 5a, Socs1−/− BMDCs were also resistant to cAMP-mediated suppression of cytokine production in the presence of LPS+IFNγ. IFNγ dose-dependently counteracted the suppressive effects of cAMP in Socs1−/− BMDCs (Fig. 5b). It is notable that IFNγ partially antagonized the suppressive effect of PGE2 and cAMP in Socs1+/+ BMDCs (Fig. 5b), although this antagonism was not observed in Stat1−/− BMDCs (Fig. 5c). These data suggest that the antagonistic effect of IFNγ/STAT1 pathway against PGE2 and cAMP was amplified in Socs1−/− BMDCs.

Bottom Line: Additionally, we find an essential role for prostaglandin E2 (PGE2) in the maintenance of tolerance within the intestine in the absence of Tregs.Socs1(-/-) dendritic cells are resistant to PGE2-mediated immunosuppression because of dysregulated cytokine signalling.Thus, we propose that SOCS1 and PGE2, potentially interacting together, act as an alternative intestinal tolerance mechanism distinct from IL-10 and Tregs.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

ABSTRACT
Interleukin 10 (IL-10) and regulatory T cells (Tregs) maintain tolerance to intestinal microorganisms. However, Il10(-/-)Rag2(-/-) mice, which lack IL-10 and Tregs, remain healthy, suggesting the existence of other mechanisms of tolerance. Here, we identify suppressor of cytokine signalling 1 (SOCS1) as an essential mediator of immune tolerance in the intestine. Socs1(-/-)Rag2(-/-) mice develop severe colitis, which can be prevented by the reduction of microbiota and the transfer of IL-10-sufficient Tregs. Additionally, we find an essential role for prostaglandin E2 (PGE2) in the maintenance of tolerance within the intestine in the absence of Tregs. Socs1(-/-) dendritic cells are resistant to PGE2-mediated immunosuppression because of dysregulated cytokine signalling. Thus, we propose that SOCS1 and PGE2, potentially interacting together, act as an alternative intestinal tolerance mechanism distinct from IL-10 and Tregs.

Show MeSH
Related in: MedlinePlus