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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 renders Socs1−/− BMDCs resistant to PGE2 but not to IL-10.(a–e, g, h) Socs1+/+ (black bars) and Socs1−/− (grey bars) BMDCs (5×105 cells per ml) induced from the bone marrow cells of Rag2−/− and Socs1−/−Rag2−/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. 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. (a) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS (10 ng ml−1) and murine IFNγ (10 ng ml−1) in the presence of graded concentrations of recombinant murine IL-10. (b) Socs1+/+ and Socs1−/− BMDCs were co-cultured with 2.5×105 cells per ml of CD4+CD25high T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with LPS+IFNγ (10 ng ml−1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml−1). (c) Socs1+/+ BMDCs were stimualted with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml−1). (d) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). **P<0.01. (e) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME. The CME filtrated through YM-3 ultrafiltration columns (Millipore), which remove molecules higher than 3,000 Da, retained a suppressive function. IL-10 was used as an experimental control. (f) Tissue extracts from the indicated organs were prepared as described in the Methods, and the levels of PGE2 were determined by ELISA. Error bars indicate +s.d. of triplicate ELISA measurements for each sample. (g) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence of PGE2 (1 nM) or graded concentrations of CME with or without PTGER4 antagonist (ONO-AE3-208; 100 nM). (h) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of PGE2. **P<0.01. Data are representative of two (f) to three (a–e, g, h) independent experiments.
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f3: IFNγ renders Socs1−/− BMDCs resistant to PGE2 but not to IL-10.(a–e, g, h) Socs1+/+ (black bars) and Socs1−/− (grey bars) BMDCs (5×105 cells per ml) induced from the bone marrow cells of Rag2−/− and Socs1−/−Rag2−/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. 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. (a) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS (10 ng ml−1) and murine IFNγ (10 ng ml−1) in the presence of graded concentrations of recombinant murine IL-10. (b) Socs1+/+ and Socs1−/− BMDCs were co-cultured with 2.5×105 cells per ml of CD4+CD25high T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with LPS+IFNγ (10 ng ml−1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml−1). (c) Socs1+/+ BMDCs were stimualted with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml−1). (d) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). **P<0.01. (e) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME. The CME filtrated through YM-3 ultrafiltration columns (Millipore), which remove molecules higher than 3,000 Da, retained a suppressive function. IL-10 was used as an experimental control. (f) Tissue extracts from the indicated organs were prepared as described in the Methods, and the levels of PGE2 were determined by ELISA. Error bars indicate +s.d. of triplicate ELISA measurements for each sample. (g) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence of PGE2 (1 nM) or graded concentrations of CME with or without PTGER4 antagonist (ONO-AE3-208; 100 nM). (h) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of PGE2. **P<0.01. Data are representative of two (f) to three (a–e, g, h) independent experiments.

Mentions: Our Treg transfer experiment into Socs1−/−Rag2−/− mice indicated that suppression by Treg-derived IL-10 could overwhelm the inflammatory effect of SOCS1 deficiency. In accordance with this scenario, IL-10 suppressed the TNFα and IL-12p70 production from Socs1−/− BMDCs stimulated with LPS or LPS+IFNγ as efficiently as from Socs1+/+ BMDCs (Fig. 3a, data for LPS alone shown in Supplementary Fig. S1a). Similarly, Tregs suppressed inflammatory cytokine production from BMDCs in an IL-10-dependent manner and this Treg-mediated suppression was also observed in Socs1−/− BMDCs (Fig. 3b). These observations suggested that IL-10/Treg-mediated suppression was not impaired by SOCS1 deficiency; therefore, we hypothesized that another immunosuppressive mechanism was altered in the colons of Socs1−/−Rag2−/− mice.


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 renders Socs1−/− BMDCs resistant to PGE2 but not to IL-10.(a–e, g, h) Socs1+/+ (black bars) and Socs1−/− (grey bars) BMDCs (5×105 cells per ml) induced from the bone marrow cells of Rag2−/− and Socs1−/−Rag2−/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. 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. (a) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS (10 ng ml−1) and murine IFNγ (10 ng ml−1) in the presence of graded concentrations of recombinant murine IL-10. (b) Socs1+/+ and Socs1−/− BMDCs were co-cultured with 2.5×105 cells per ml of CD4+CD25high T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with LPS+IFNγ (10 ng ml−1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml−1). (c) Socs1+/+ BMDCs were stimualted with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml−1). (d) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). **P<0.01. (e) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME. The CME filtrated through YM-3 ultrafiltration columns (Millipore), which remove molecules higher than 3,000 Da, retained a suppressive function. IL-10 was used as an experimental control. (f) Tissue extracts from the indicated organs were prepared as described in the Methods, and the levels of PGE2 were determined by ELISA. Error bars indicate +s.d. of triplicate ELISA measurements for each sample. (g) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence of PGE2 (1 nM) or graded concentrations of CME with or without PTGER4 antagonist (ONO-AE3-208; 100 nM). (h) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of PGE2. **P<0.01. Data are representative of two (f) to three (a–e, g, h) independent experiments.
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f3: IFNγ renders Socs1−/− BMDCs resistant to PGE2 but not to IL-10.(a–e, g, h) Socs1+/+ (black bars) and Socs1−/− (grey bars) BMDCs (5×105 cells per ml) induced from the bone marrow cells of Rag2−/− and Socs1−/−Rag2−/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. 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. (a) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS (10 ng ml−1) and murine IFNγ (10 ng ml−1) in the presence of graded concentrations of recombinant murine IL-10. (b) Socs1+/+ and Socs1−/− BMDCs were co-cultured with 2.5×105 cells per ml of CD4+CD25high T cells in the presence of soluble anti-CD3 Ab (1 μg ml−1). Cells were stimulated with LPS+IFNγ (10 ng ml−1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml−1). (c) Socs1+/+ BMDCs were stimualted with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml−1). (d) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). **P<0.01. (e) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence or absence of graded concentrations of CME. The CME filtrated through YM-3 ultrafiltration columns (Millipore), which remove molecules higher than 3,000 Da, retained a suppressive function. IL-10 was used as an experimental control. (f) Tissue extracts from the indicated organs were prepared as described in the Methods, and the levels of PGE2 were determined by ELISA. Error bars indicate +s.d. of triplicate ELISA measurements for each sample. (g) Socs1+/+ BMDCs were stimulated with LPS (10 ng ml−1) in the presence of PGE2 (1 nM) or graded concentrations of CME with or without PTGER4 antagonist (ONO-AE3-208; 100 nM). (h) Socs1+/+ and Socs1−/− BMDCs were stimulated with LPS+IFNγ (10 ng ml−1 each) in the presence of graded concentrations of PGE2. **P<0.01. Data are representative of two (f) to three (a–e, g, h) independent experiments.
Mentions: Our Treg transfer experiment into Socs1−/−Rag2−/− mice indicated that suppression by Treg-derived IL-10 could overwhelm the inflammatory effect of SOCS1 deficiency. In accordance with this scenario, IL-10 suppressed the TNFα and IL-12p70 production from Socs1−/− BMDCs stimulated with LPS or LPS+IFNγ as efficiently as from Socs1+/+ BMDCs (Fig. 3a, data for LPS alone shown in Supplementary Fig. S1a). Similarly, Tregs suppressed inflammatory cytokine production from BMDCs in an IL-10-dependent manner and this Treg-mediated suppression was also observed in Socs1−/− BMDCs (Fig. 3b). These observations suggested that IL-10/Treg-mediated suppression was not impaired by SOCS1 deficiency; therefore, we hypothesized that another immunosuppressive mechanism was altered in the colons of Socs1−/−Rag2−/− mice.

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