Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1

Magdalena Niedzielska, Faizal A. M. Raffi, Jurjen Tel, Sandra Muench, Katrin Jozefowski, Nour Alati, Katharina Lahl, Jorg Mages, Ulrike Billmeier, Matthias Schiemann, Uwe K. Appelt, Stefan Wirtz, Tim Sparwasser, Hubertus Hochrein, Carl G. Figdor, Stephen M. Keyse, Roland Lang (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    Plasmacytoid dendritic cells (pDCs) efficiently produce large amounts of type I IFN in response to TLR7 and TLR9 ligands, whereas conventional DCs (cDCs) predominantly secrete high levels of the cytokines IL-10 and IL-12. The molecular basis underlying this distinct phenotype is not well understood. In this study, we identified the MAPK phosphatase Dusp9/MKP-4 by transcriptome analysis as selectively expressed in pDCs, but not cDCs. We confirmed the constitutive expression of Dusp9 at the protein level in pDCs generated in vitro by culture with Flt3 ligand and ex vivo in sorted splenic pDCs. Dusp9 expression was low in B220 bone marrow precursors and was upregulated during pDC differentiation, concomitant with established pDC markers. Higher expression of Dusp9 in pDCs correlated with impaired phosphorylation of the MAPK ERK1/2 upon TLR9 stimulation. Notably, Dusp9 was not expressed at detectable levels in human pDCs, although these displayed similarly impaired activation of ERK1/2 MAPK compared with cDCs. Enforced retroviral expression of Dusp9 in mouse GM-CSF–induced cDCs increased the expression of TLR9-induced IL-12p40 and IFN-β, but not of IL-10. Conditional deletion of Dusp9 in pDCs was effectively achieved in Dusp9flox/flox; CD11c-Cre mice at the mRNA and protein levels. However, the lack of Dusp9 in pDC did not restore ERK1/2 activation after TLR9 stimulation and only weakly affected IFN-β and IL-12p40 production. Taken together, our results suggest that expression of Dusp9 is sufficient to impair ERK1/2 activation and enhance IFN-β expression. However, despite selective expression in pDCs, Dusp9 is not essential for high-level IFN-β production by these cells.
    Original languageEnglish
    Pages (from-to)1753-1762
    Number of pages10
    JournalJournal of Immunology
    Volume195
    Issue number4
    Early online date13 Jul 2015
    DOIs
    Publication statusPublished - 15 Aug 2015

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    Dual Specificity Phosphatase 1
    Dendritic Cells
    Interleukin-12 Subunit p40
    Interleukin-10
    Gene Expression Profiling
    Interleukin-12
    Proteins
    Bone Marrow
    Phosphorylation
    Cytokines
    Ligands
    Phenotype
    Messenger RNA

    Cite this

    Niedzielska, Magdalena ; Raffi, Faizal A. M. ; Tel, Jurjen ; Muench, Sandra ; Jozefowski, Katrin ; Alati, Nour ; Lahl, Katharina ; Mages, Jorg ; Billmeier, Ulrike ; Schiemann, Matthias ; Appelt, Uwe K. ; Wirtz, Stefan ; Sparwasser, Tim ; Hochrein, Hubertus ; Figdor, Carl G. ; Keyse, Stephen M. ; Lang, Roland. / Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1. In: Journal of Immunology. 2015 ; Vol. 195, No. 4. pp. 1753-1762.
    @article{defef51a447b46f880fc6737fcfa1052,
    title = "Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1",
    abstract = "Plasmacytoid dendritic cells (pDCs) efficiently produce large amounts of type I IFN in response to TLR7 and TLR9 ligands, whereas conventional DCs (cDCs) predominantly secrete high levels of the cytokines IL-10 and IL-12. The molecular basis underlying this distinct phenotype is not well understood. In this study, we identified the MAPK phosphatase Dusp9/MKP-4 by transcriptome analysis as selectively expressed in pDCs, but not cDCs. We confirmed the constitutive expression of Dusp9 at the protein level in pDCs generated in vitro by culture with Flt3 ligand and ex vivo in sorted splenic pDCs. Dusp9 expression was low in B220− bone marrow precursors and was upregulated during pDC differentiation, concomitant with established pDC markers. Higher expression of Dusp9 in pDCs correlated with impaired phosphorylation of the MAPK ERK1/2 upon TLR9 stimulation. Notably, Dusp9 was not expressed at detectable levels in human pDCs, although these displayed similarly impaired activation of ERK1/2 MAPK compared with cDCs. Enforced retroviral expression of Dusp9 in mouse GM-CSF–induced cDCs increased the expression of TLR9-induced IL-12p40 and IFN-β, but not of IL-10. Conditional deletion of Dusp9 in pDCs was effectively achieved in Dusp9flox/flox; CD11c-Cre mice at the mRNA and protein levels. However, the lack of Dusp9 in pDC did not restore ERK1/2 activation after TLR9 stimulation and only weakly affected IFN-β and IL-12p40 production. Taken together, our results suggest that expression of Dusp9 is sufficient to impair ERK1/2 activation and enhance IFN-β expression. However, despite selective expression in pDCs, Dusp9 is not essential for high-level IFN-β production by these cells.",
    author = "Magdalena Niedzielska and Raffi, {Faizal A. M.} and Jurjen Tel and Sandra Muench and Katrin Jozefowski and Nour Alati and Katharina Lahl and Jorg Mages and Ulrike Billmeier and Matthias Schiemann and Appelt, {Uwe K.} and Stefan Wirtz and Tim Sparwasser and Hubertus Hochrein and Figdor, {Carl G.} and Keyse, {Stephen M.} and Roland Lang",
    year = "2015",
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    doi = "10.4049/?jimmunol.1400658",
    language = "English",
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    Niedzielska, M, Raffi, FAM, Tel, J, Muench, S, Jozefowski, K, Alati, N, Lahl, K, Mages, J, Billmeier, U, Schiemann, M, Appelt, UK, Wirtz, S, Sparwasser, T, Hochrein, H, Figdor, CG, Keyse, SM & Lang, R 2015, 'Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1', Journal of Immunology, vol. 195, no. 4, pp. 1753-1762. https://doi.org/10.4049/?jimmunol.1400658

    Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1. / Niedzielska, Magdalena; Raffi, Faizal A. M.; Tel, Jurjen; Muench, Sandra; Jozefowski, Katrin; Alati, Nour; Lahl, Katharina; Mages, Jorg; Billmeier, Ulrike; Schiemann, Matthias; Appelt, Uwe K.; Wirtz, Stefan; Sparwasser, Tim; Hochrein, Hubertus; Figdor, Carl G.; Keyse, Stephen M.; Lang, Roland (Lead / Corresponding author).

    In: Journal of Immunology, Vol. 195, No. 4, 15.08.2015, p. 1753-1762.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Selective expression of the MAPK phosphatase Dusp9/MKP-4 in mouse plasmacytoid dendritic cells and regulation of IFNβ production1

    AU - Niedzielska, Magdalena

    AU - Raffi, Faizal A. M.

    AU - Tel, Jurjen

    AU - Muench, Sandra

    AU - Jozefowski, Katrin

    AU - Alati, Nour

    AU - Lahl, Katharina

    AU - Mages, Jorg

    AU - Billmeier, Ulrike

    AU - Schiemann, Matthias

    AU - Appelt, Uwe K.

    AU - Wirtz, Stefan

    AU - Sparwasser, Tim

    AU - Hochrein, Hubertus

    AU - Figdor, Carl G.

    AU - Keyse, Stephen M.

    AU - Lang, Roland

    PY - 2015/8/15

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    N2 - Plasmacytoid dendritic cells (pDCs) efficiently produce large amounts of type I IFN in response to TLR7 and TLR9 ligands, whereas conventional DCs (cDCs) predominantly secrete high levels of the cytokines IL-10 and IL-12. The molecular basis underlying this distinct phenotype is not well understood. In this study, we identified the MAPK phosphatase Dusp9/MKP-4 by transcriptome analysis as selectively expressed in pDCs, but not cDCs. We confirmed the constitutive expression of Dusp9 at the protein level in pDCs generated in vitro by culture with Flt3 ligand and ex vivo in sorted splenic pDCs. Dusp9 expression was low in B220− bone marrow precursors and was upregulated during pDC differentiation, concomitant with established pDC markers. Higher expression of Dusp9 in pDCs correlated with impaired phosphorylation of the MAPK ERK1/2 upon TLR9 stimulation. Notably, Dusp9 was not expressed at detectable levels in human pDCs, although these displayed similarly impaired activation of ERK1/2 MAPK compared with cDCs. Enforced retroviral expression of Dusp9 in mouse GM-CSF–induced cDCs increased the expression of TLR9-induced IL-12p40 and IFN-β, but not of IL-10. Conditional deletion of Dusp9 in pDCs was effectively achieved in Dusp9flox/flox; CD11c-Cre mice at the mRNA and protein levels. However, the lack of Dusp9 in pDC did not restore ERK1/2 activation after TLR9 stimulation and only weakly affected IFN-β and IL-12p40 production. Taken together, our results suggest that expression of Dusp9 is sufficient to impair ERK1/2 activation and enhance IFN-β expression. However, despite selective expression in pDCs, Dusp9 is not essential for high-level IFN-β production by these cells.

    AB - Plasmacytoid dendritic cells (pDCs) efficiently produce large amounts of type I IFN in response to TLR7 and TLR9 ligands, whereas conventional DCs (cDCs) predominantly secrete high levels of the cytokines IL-10 and IL-12. The molecular basis underlying this distinct phenotype is not well understood. In this study, we identified the MAPK phosphatase Dusp9/MKP-4 by transcriptome analysis as selectively expressed in pDCs, but not cDCs. We confirmed the constitutive expression of Dusp9 at the protein level in pDCs generated in vitro by culture with Flt3 ligand and ex vivo in sorted splenic pDCs. Dusp9 expression was low in B220− bone marrow precursors and was upregulated during pDC differentiation, concomitant with established pDC markers. Higher expression of Dusp9 in pDCs correlated with impaired phosphorylation of the MAPK ERK1/2 upon TLR9 stimulation. Notably, Dusp9 was not expressed at detectable levels in human pDCs, although these displayed similarly impaired activation of ERK1/2 MAPK compared with cDCs. Enforced retroviral expression of Dusp9 in mouse GM-CSF–induced cDCs increased the expression of TLR9-induced IL-12p40 and IFN-β, but not of IL-10. Conditional deletion of Dusp9 in pDCs was effectively achieved in Dusp9flox/flox; CD11c-Cre mice at the mRNA and protein levels. However, the lack of Dusp9 in pDC did not restore ERK1/2 activation after TLR9 stimulation and only weakly affected IFN-β and IL-12p40 production. Taken together, our results suggest that expression of Dusp9 is sufficient to impair ERK1/2 activation and enhance IFN-β expression. However, despite selective expression in pDCs, Dusp9 is not essential for high-level IFN-β production by these cells.

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    DO - 10.4049/?jimmunol.1400658

    M3 - Article

    VL - 195

    SP - 1753

    EP - 1762

    JO - Journal of Immunology

    JF - Journal of Immunology

    SN - 0022-1767

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