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TLR2/6 and TLR4-activated macrophages contribute to islet inflammation and impair beta cell insulin gene expression via IL-1 and IL-6

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Abstract

Aims/hypothesis

Inflammation contributes to pancreatic beta cell dysfunction in type 2 diabetes. Toll-like receptor (TLR)-2 and -4 ligands are increased systemically in recently diagnosed type 2 diabetes patients, and TLR2- and TLR4-deficient mice are protected from the metabolic consequences of a high-fat diet. Here we investigated the role of macrophages in TLR2/6- and TLR4-mediated effects on islet inflammation and beta cell function.

Methods

Genetic and pharmacological approaches were used to determine the effects of TLR2/6 and TLR4 ligands on mouse islets, human islets and purified rat beta cells. Islet macrophages were depleted and sorted by flow cytometry and the effects of TLR2/6- and TLR4-activated bone-marrow-derived macrophages (BMDMs) on beta cell function were assessed.

Results

Macrophages contributed to TLR2/6- and TLR4-induced islet Il1a/IL1A and Il1b/IL1B mRNA expression in mouse and human islets and IL-1β secretion from human islets. TLR2/6 and TLR4 ligands also reduced insulin gene expression; however, this occurred in a non-beta cell autonomous manner. TLR2/6- and TLR4-activated BMDMs reduced beta cell insulin secretion partly via reducing Ins1, Ins2, and Pdx1 mRNA expression. Antagonism of the IL-1 receptor and neutralisation of IL-6 completely reversed the effects of activated macrophages on beta cell gene expression.

Conclusions/interpretation

We conclude that islet macrophages are major contributors to islet IL-1β secretion in response to TLR2/6 and TLR4 ligands. BMDMs stimulated with TLR2/6 and TLR4 ligands reduce insulin secretion from pancreatic beta cells, partly via IL-1β- and IL-6-mediated decreased insulin gene expression.

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Abbreviations

BMDM:

Bone-marrow-derived macrophage

CCL2:

Chemokine (C-C motif) ligand 2

DAMPs:

Danger-associated molecular patterns

GSIS:

Glucose-stimulated insulin secretion

LPS:

Lipopolysaccharide

PAMPs:

Pathogen-associated molecular patterns

qPCR:

Quantitative PCR

TLR:

Toll-like receptor

WT:

Wild-type

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Acknowledgements

We thank L. Xu and M. Komba for technical assistance provided by the CFRI FACS core and islet isolation core facilities, respectively.

Funding

This work was supported by funding from the Child and Family Research Institute (JAE), the University of British Columbia (JAE), the Canadian Institutes of Health Research (PCN-110793 and PNI-120292; JAE), the European Research Council (KM), the Diabetes Competence Network KKNDm supported by the Federal Ministry of Germany (BMBF; KM), and a collaborative research agreement with Servier (JAE, KM). JAE has salary support from a Child and Family Research Institute Investigator Award and a Canadian Diabetes Association scholar award. DN is supported by a UBC Transplantation Training Program and a CIHR-Vanier Canada Graduate Scholarship. CW-R is supported by a CIHR-Vanier Canada Graduate Scholarship.

Duality of interest

CS-K and BG are employees of Servier, France. All other authors declare that there is no duality of interest associated with their contribution to this manuscript.

Contribution statement

DN, MD, WH, RK, AS, SR, CW-R, AC, MS designed and performed experiments, and analysed data. KM designed experiments and CS-K and BG contributed to the conception and design of the study. All authors edited the manuscript and approved the final version. JAE supervised the study, designed and performed experiments, analysed data and wrote the manuscript. JAE is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

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Authors and Affiliations

  1. Department of Surgery, Faculty of Medicine, The University of British Columbia, Child and Family Research Institute, 950 W 28th Ave, Vancouver, BC, Canada, V5Z 4H4

    Dominika Nackiewicz, Meixia Dan, Rosa Kim, Anisa Salmi, Amanda Cunningham, Madeleine Speck & Jan A. Ehses

  2. Center for Biomolecular Interactions, The University of Bremen, Bremen, Germany

    Wei He & Kathrin Maedler

  3. Department of Genetic Medicine and Development, University Medical Center, University of Geneva, Geneva, Switzerland

    Sabine Rütti

  4. Department of Pathology and Laboratory Medicine, Faculty of Medicine, The University of British Columbia, Child and Family Research Institute, Vancouver, BC, Canada

    Clara Westwell-Roper

  5. ADIR – Groupe de Recherche Servier, Suresnes, France

    Carole Schuster-Klein & Beatrice Guardiola

  6. Department of Cellular and Physiological Sciences, Faculty of Medicine, The University of British Columbia, Child and Family Research Institute, Vancouver, BC, Canada

    Jan A. Ehses

Authors
  1. Dominika Nackiewicz
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  2. Meixia Dan
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  3. Wei He
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  4. Rosa Kim
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  12. Kathrin Maedler
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  13. Jan A. Ehses
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Corresponding author

Correspondence to Jan A. Ehses.

Additional information

Dominika Nackiewicz and Meixia Dan contributed equally to this study.

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Nackiewicz, D., Dan, M., He, W. et al. TLR2/6 and TLR4-activated macrophages contribute to islet inflammation and impair beta cell insulin gene expression via IL-1 and IL-6. Diabetologia 57, 1645–1654 (2014). https://doi.org/10.1007/s00125-014-3249-1

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