Differential control of Mincle-dependent cord factor recognition and macrophage responses by the transcription factors C/EBPβ and HIF1α

Hanne Schoenen; Alexandra Huber; Nada Sonda; Stephanie Zimmermann; Jonathan Jantsch; Bernd Lepenies; Vincenzo Bronte; Roland Lang

doi:10.4049/jimmunol.1301593

Differential control of Mincle-dependent cord factor recognition and macrophage responses by the transcription factors C/EBPβ and HIF1α

J Immunol. 2014 Oct 1;193(7):3664-75. doi: 10.4049/jimmunol.1301593. Epub 2014 Aug 25.

Authors

Hanne Schoenen¹, Alexandra Huber¹, Nada Sonda², Stephanie Zimmermann³, Jonathan Jantsch⁴, Bernd Lepenies³, Vincenzo Bronte⁵, Roland Lang⁶

Affiliations

¹ Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Friedrich Alexander Universität Erlangen-Nürnberg, 91054 Erlangen, Germany;
² Oncology and Immunology Section, Department of Surgery, Oncology and Gastroenterology, University of Padua, 35128, Padua, Italy;
³ Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14476 Potsdam, Germany; Freie Universität Berlin, Institute of Chemistry and Biochemistry, 14195 Berlin, Germany;
⁴ Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Friedrich Alexander Universität Erlangen-Nürnberg, 91054 Erlangen, Germany; Institut für Klinische Mikrobiologie und Hygiene, Universitätsklinikum Regensburg, 93053 Regensburg, Germany; and.
⁵ Immunology Section, Department of Pathology, Verona University Hospital, University of Verona, 37134 Verona, Italy.
⁶ Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Friedrich Alexander Universität Erlangen-Nürnberg, 91054 Erlangen, Germany; [email protected].

PMID: 25156364
DOI: 10.4049/jimmunol.1301593

Abstract

Trehalose-6,6-dimycolate (TDM), the mycobacterial cord factor, and its synthetic analog Trehalose-6,6-dibehenate (TDB) bind to the C-type lectin receptors macrophage-inducible C-type lectin (Mincle) and Mcl to activate macrophages. Genetically, the transcriptional response to TDB/TDM has been defined to require FcRγ-Syk-Card9 signaling. However, TDB/TDM-triggered kinase activation has not been studied well, and it is largely unknown which transcriptional regulators bring about inflammatory gene expression. In this article, we report that TDB/TDM caused only weak Syk-phosphorylation in resting macrophages, consistent with low basal Mincle expression. However, LPS-priming caused MYD88-dependent upregulation of Mincle, resulting in enhanced TDB/TDM-induced kinase activation and more rapid inflammatory gene expression. TLR-induced Mincle expression partially circumvented the requirement for Mcl in the response to TDB/TDM. To dissect transcriptional responses to TDB/TDM, we mined microarray data and identified early growth response (Egr) family transcription factors as direct Mincle target genes, whereas upregulation of Cebpb and Hif1a required new protein synthesis. Macrophages and dendritic cells lacking C/EBPβ showed nearly complete abrogation of TDB/TDM responsiveness, but also failed to upregulate Mincle. Retroviral rescue of Mincle expression in Cebpb-deficient cells restored induction of Egr1, but not of G-CSF. This pattern of C/EBPβ dependence was also observed after stimulation with the Dectin-1 ligand Curdlan. Inducible expression of hypoxia-inducible factor 1α (HIF1α) also required C/EBPβ. In turn, HIF1α was not required for Mincle expression, kinase activation, and Egr1 or Csf3 expression, but critically contributed to NO production. Taken together, we identify C/EBPβ as central hub in Mincle expression and inflammatory gene induction, whereas HIF1α controls Nos2 expression. C/EBPβ also connects TLR signals to cord factor responsiveness through MYD88-dependent upregulation of Mincle.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CCAAT-Enhancer-Binding Protein-beta / genetics
CCAAT-Enhancer-Binding Protein-beta / immunology*
Cord Factors / chemistry
Cord Factors / pharmacology*
Dendritic Cells / cytology
Dendritic Cells / immunology
Early Growth Response Protein 1 / genetics
Early Growth Response Protein 1 / immunology
Enzyme Activation / drug effects
Enzyme Activation / genetics
Enzyme Activation / immunology
Granulocyte Colony-Stimulating Factor / immunology
Hypoxia-Inducible Factor 1, alpha Subunit / genetics
Hypoxia-Inducible Factor 1, alpha Subunit / immunology*
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / immunology
Lectins, C-Type / genetics
Lectins, C-Type / immunology*
Macrophages / cytology
Macrophages / immunology*
Membrane Proteins / genetics
Membrane Proteins / immunology*
Mice
Mice, Knockout
Mycobacterium tuberculosis / chemistry*
Myeloid Differentiation Factor 88 / genetics
Myeloid Differentiation Factor 88 / immunology
Nitric Oxide / genetics
Nitric Oxide / immunology
Nitric Oxide Synthase Type II / genetics
Nitric Oxide Synthase Type II / immunology
Phosphorylation / drug effects
Phosphorylation / genetics
Phosphorylation / immunology
Protein Biosynthesis / drug effects
Protein Biosynthesis / genetics
Protein Biosynthesis / immunology
Protein-Tyrosine Kinases / genetics
Protein-Tyrosine Kinases / immunology
Syk Kinase
Toll-Like Receptors / genetics
Toll-Like Receptors / immunology
Up-Regulation / drug effects*
Up-Regulation / genetics
Up-Regulation / immunology
beta-Glucans / chemistry
beta-Glucans / pharmacology

Substances

CCAAT-Enhancer-Binding Protein-beta
Cebpb protein, mouse
Clecsf8 protein, mouse
Cord Factors
Early Growth Response Protein 1
Egr1 protein, mouse
Hif1a protein, mouse
Hypoxia-Inducible Factor 1, alpha Subunit
Intracellular Signaling Peptides and Proteins
Lectins, C-Type
Membrane Proteins
Myd88 protein, mouse
Myeloid Differentiation Factor 88
Toll-Like Receptors
beta-Glucans
dectin 1
Granulocyte Colony-Stimulating Factor
Nitric Oxide
curdlan
Nitric Oxide Synthase Type II
Nos2 protein, mouse
Protein-Tyrosine Kinases
Syk Kinase
Syk protein, mouse