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Dual-specificity phosphatase 6 regulates CD4+ T-cell functions and restrains spontaneous colitis in IL-10-deficient mice.

Bertin S, Lozano-Ruiz B, Bachiller V, García-Martínez I, Herdman S, Zapater P, Francés R, Such J, Lee J, Raz E, González-Navajas JM - Mucosal Immunol (2014)

Bottom Line: DUSP6 preferentially dephosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1/2) rendering them inactive.In summary, DUSP6 regulates CD4(+) T-cell activation and differentiation by inhibiting the TCR-dependent ERK1/2 activation.DUSP6 might therefore be a potential intervention target for limiting aberrant T-cell responses in T-cell-mediated diseases, such as inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Allergy and Immunology, University of California San Diego, La Jolla, California, USA.

ABSTRACT
Mitogen-activated protein kinase (MAPK) phosphatases are dual-specificity phosphatases (DUSPs) that dephosphorylate phosphothreonine and phosphotyrosine residues within MAPKs. DUSP6 preferentially dephosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1/2) rendering them inactive. Here, we study the role of DUSP6 in CD4(+) T-cell function, differentiation, and inflammatory profile in the colon. Upon T-cell receptor (TCR) stimulation, DUSP6 knockout (Dusp6(-/-)) CD4(+) T cells showed increased ERK1/2 activation, proliferation, T helper 1 differentiation, and interferon-γ production, as well as a marked decrease in survival, interleukin- 17A (IL-17A) secretion, and regulatory T-cell function. To analyze the role of DUSP6 in vivo, we employed the Il10(-/-) model of colitis and generated Il10(-/-)/Dusp6(-/-) double-knockout mice. Il10(-/-)/Dusp6(-/-) mice suffered from accelerated and exacerbated spontaneous colitis, which was prevented by ERK1/2 inhibition. ERK1/2 inhibition also augmented regulatory T-cell differentiation in vitro and in vivo in both C57Bl/6 and Dusp6(-/-) mice. In summary, DUSP6 regulates CD4(+) T-cell activation and differentiation by inhibiting the TCR-dependent ERK1/2 activation. DUSP6 might therefore be a potential intervention target for limiting aberrant T-cell responses in T-cell-mediated diseases, such as inflammatory bowel disease.

No MeSH data available.


Related in: MedlinePlus

DUSP6 deficiency facilitates Th1 differentiation in vitro(a) and (c) Flow cytometry analysis of intracellular cytokines in naïve CD4+ T cells from WT and Dusp6−/− mice cultured in Th1 (a) or Th17 (c) polarizing conditions for five days, and re-stimulated with anti-CD3/28 antibodies for 6 hours. (b) and (d) Cytokine levels measured by ELISA in supernatants of naïve T cells cultured in Th1 (b) or Th17 (d) conditions and re-stimulated with anti-CD3/28 antibodies for 48 hours. Data in this figure represent pooled results from four independent experiments. Error bars represent standard deviation. **P<0.01
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Figure 3: DUSP6 deficiency facilitates Th1 differentiation in vitro(a) and (c) Flow cytometry analysis of intracellular cytokines in naïve CD4+ T cells from WT and Dusp6−/− mice cultured in Th1 (a) or Th17 (c) polarizing conditions for five days, and re-stimulated with anti-CD3/28 antibodies for 6 hours. (b) and (d) Cytokine levels measured by ELISA in supernatants of naïve T cells cultured in Th1 (b) or Th17 (d) conditions and re-stimulated with anti-CD3/28 antibodies for 48 hours. Data in this figure represent pooled results from four independent experiments. Error bars represent standard deviation. **P<0.01

Mentions: Our previous observations showing a different production of IFN-γ and IL-17A by Dusp6−/− CD4+ T cells (Figure 1 and Supplementary Figure S1) suggested that DUSP6 might be involved in the differentiation of Th cell subsets. To test this hypothesis, we isolated naïve CD4+ T cells from the spleen of WT and Dusp6−/− mice by FACS sorting, and cultured them under Th1 or Th17 polarizing conditions. After 5 days of differentiation in vitro, we re-stimulated the cells with anti-CD3/CD28 antibodies and analyzed IFN-γ and IL-17A production by FACS intracellular staining and ELISA. Under Th1 conditions, the number of IFN-γ producing cells was increased in Dusp6−/− CD4+ T cells when compared to WT T cells (Figure 3a). Consistent with these data, the protein level of IFN-γ was also increased in culture supernatants of Dusp6−/− CD4+ T cells (Figure 3b). By contrast, under Th17 polarizing conditions, we found that both the number of IL-17A producing cells and the level of IL-17A in culture supernatants were diminished in Dusp6−/− CD4+ T cells when compared to WT control cells (Figure 3c and 3d). These data suggest that DUSP6 regulates the development of CD4+ T cell subsets by inhibiting Th1 differentiation and favoring Th17 differentiation.


Dual-specificity phosphatase 6 regulates CD4+ T-cell functions and restrains spontaneous colitis in IL-10-deficient mice.

Bertin S, Lozano-Ruiz B, Bachiller V, García-Martínez I, Herdman S, Zapater P, Francés R, Such J, Lee J, Raz E, González-Navajas JM - Mucosal Immunol (2014)

DUSP6 deficiency facilitates Th1 differentiation in vitro(a) and (c) Flow cytometry analysis of intracellular cytokines in naïve CD4+ T cells from WT and Dusp6−/− mice cultured in Th1 (a) or Th17 (c) polarizing conditions for five days, and re-stimulated with anti-CD3/28 antibodies for 6 hours. (b) and (d) Cytokine levels measured by ELISA in supernatants of naïve T cells cultured in Th1 (b) or Th17 (d) conditions and re-stimulated with anti-CD3/28 antibodies for 48 hours. Data in this figure represent pooled results from four independent experiments. Error bars represent standard deviation. **P<0.01
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4363301&req=5

Figure 3: DUSP6 deficiency facilitates Th1 differentiation in vitro(a) and (c) Flow cytometry analysis of intracellular cytokines in naïve CD4+ T cells from WT and Dusp6−/− mice cultured in Th1 (a) or Th17 (c) polarizing conditions for five days, and re-stimulated with anti-CD3/28 antibodies for 6 hours. (b) and (d) Cytokine levels measured by ELISA in supernatants of naïve T cells cultured in Th1 (b) or Th17 (d) conditions and re-stimulated with anti-CD3/28 antibodies for 48 hours. Data in this figure represent pooled results from four independent experiments. Error bars represent standard deviation. **P<0.01
Mentions: Our previous observations showing a different production of IFN-γ and IL-17A by Dusp6−/− CD4+ T cells (Figure 1 and Supplementary Figure S1) suggested that DUSP6 might be involved in the differentiation of Th cell subsets. To test this hypothesis, we isolated naïve CD4+ T cells from the spleen of WT and Dusp6−/− mice by FACS sorting, and cultured them under Th1 or Th17 polarizing conditions. After 5 days of differentiation in vitro, we re-stimulated the cells with anti-CD3/CD28 antibodies and analyzed IFN-γ and IL-17A production by FACS intracellular staining and ELISA. Under Th1 conditions, the number of IFN-γ producing cells was increased in Dusp6−/− CD4+ T cells when compared to WT T cells (Figure 3a). Consistent with these data, the protein level of IFN-γ was also increased in culture supernatants of Dusp6−/− CD4+ T cells (Figure 3b). By contrast, under Th17 polarizing conditions, we found that both the number of IL-17A producing cells and the level of IL-17A in culture supernatants were diminished in Dusp6−/− CD4+ T cells when compared to WT control cells (Figure 3c and 3d). These data suggest that DUSP6 regulates the development of CD4+ T cell subsets by inhibiting Th1 differentiation and favoring Th17 differentiation.

Bottom Line: DUSP6 preferentially dephosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1/2) rendering them inactive.In summary, DUSP6 regulates CD4(+) T-cell activation and differentiation by inhibiting the TCR-dependent ERK1/2 activation.DUSP6 might therefore be a potential intervention target for limiting aberrant T-cell responses in T-cell-mediated diseases, such as inflammatory bowel disease.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology, Allergy and Immunology, University of California San Diego, La Jolla, California, USA.

ABSTRACT
Mitogen-activated protein kinase (MAPK) phosphatases are dual-specificity phosphatases (DUSPs) that dephosphorylate phosphothreonine and phosphotyrosine residues within MAPKs. DUSP6 preferentially dephosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1/2) rendering them inactive. Here, we study the role of DUSP6 in CD4(+) T-cell function, differentiation, and inflammatory profile in the colon. Upon T-cell receptor (TCR) stimulation, DUSP6 knockout (Dusp6(-/-)) CD4(+) T cells showed increased ERK1/2 activation, proliferation, T helper 1 differentiation, and interferon-γ production, as well as a marked decrease in survival, interleukin- 17A (IL-17A) secretion, and regulatory T-cell function. To analyze the role of DUSP6 in vivo, we employed the Il10(-/-) model of colitis and generated Il10(-/-)/Dusp6(-/-) double-knockout mice. Il10(-/-)/Dusp6(-/-) mice suffered from accelerated and exacerbated spontaneous colitis, which was prevented by ERK1/2 inhibition. ERK1/2 inhibition also augmented regulatory T-cell differentiation in vitro and in vivo in both C57Bl/6 and Dusp6(-/-) mice. In summary, DUSP6 regulates CD4(+) T-cell activation and differentiation by inhibiting the TCR-dependent ERK1/2 activation. DUSP6 might therefore be a potential intervention target for limiting aberrant T-cell responses in T-cell-mediated diseases, such as inflammatory bowel disease.

No MeSH data available.


Related in: MedlinePlus