Advertisement

Diabetologia

, Volume 57, Issue 8, pp 1645–1654 | Cite as

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

  • Dominika Nackiewicz
  • Meixia Dan
  • Wei He
  • Rosa Kim
  • Anisa Salmi
  • Sabine Rütti
  • Clara Westwell-Roper
  • Amanda Cunningham
  • Madeleine Speck
  • Carole Schuster-Klein
  • Beatrice Guardiola
  • Kathrin Maedler
  • Jan A. Ehses
Article

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.

Keywords

Beta cell Diabetes Inflammation Pancreatic islet Toll-like receptor 2 Toll-like receptor 4 

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

Notes

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.

Supplementary material

125_2014_3249_MOESM1_ESM.pdf (921 kb)
ESM Fig. 1 (PDF 921 kb)
125_2014_3249_MOESM2_ESM.pdf (2.9 mb)
ESM Fig. 2 (PDF 2,936 kb)
125_2014_3249_MOESM3_ESM.pdf (1.1 mb)
ESM Fig. 3 (PDF 1,156 kb)
125_2014_3249_MOESM4_ESM.pdf (1004 kb)
ESM Fig. 4 (PDF 1,003 kb)
125_2014_3249_MOESM5_ESM.pdf (3.3 mb)
ESM Fig. 5 (PDF 3,425 kb)

References

  1. 1.
    Wellen KE, Hotamisligil GS (2005) Inflammation, stress, and diabetes. J Clin Invest 115:1111–1119PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Pickup JC, Crook MA (1998) Is type II diabetes mellitus a disease of the innate immune system? Diabetologia 41:1241–1248PubMedCrossRefGoogle Scholar
  3. 3.
    Donath MY, Shoelson SE (2011) Type 2 diabetes as an inflammatory disease. Nat Rev Immunol 11:98–107PubMedCrossRefGoogle Scholar
  4. 4.
    Ehses JA, Perren A, Eppler E et al (2007) Increased number of islet-associated macrophages in type 2 diabetes. Diabetes 56:2356–2370PubMedCrossRefGoogle Scholar
  5. 5.
    Richardson SJ, Willcox A, Bone AJ, Foulis AK, Morgan NG (2009) Islet-associated macrophages in type 2 diabetes. Diabetologia 52:1686–1688PubMedCrossRefGoogle Scholar
  6. 6.
    Ehses JA, Lacraz G, Giroix MH et al (2009) IL-1 antagonism reduces hyperglycemia and tissue inflammation in the type 2 diabetic GK rat. Proc Natl Acad Sci U S A 106:13998–14003PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Jourdan T, Godlewski G, Cinar R et al (2013) Activation of the Nlrp3 inflammasome in infiltrating macrophages by endocannabinoids mediates beta cell loss in type 2 diabetes. Nat Med 19:1132–1140PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Eguchi K, Manabe I, Oishi-Tanaka Y et al (2012) Saturated fatty acid and TLR signaling link beta cell dysfunction and islet inflammation. Cell Metab 15:518–533PubMedCrossRefGoogle Scholar
  9. 9.
    Homo-Delarche F, Calderari S, Irminger JC et al (2006) Islet inflammation and fibrosis in a spontaneous model of type 2 diabetes, the GK rat. Diabetes 55:1625–1633PubMedCrossRefGoogle Scholar
  10. 10.
    Ehses JA, Meier DT, Wueest S et al (2010) Toll-like receptor 2-deficient mice are protected from insulin resistance and beta cell dysfunction induced by a high-fat diet. Diabetologia 53:1795–1806PubMedCrossRefGoogle Scholar
  11. 11.
    Dasu MR, Devaraj S, Park S, Jialal I (2010) Increased toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabetes Care 33:861–868PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Westwell-Roper C, Nackiewicz D, Dan M, Ehses JA (2014) Toll-like receptors and NLRP3 as central regulators of pancreatic islet inflammation in type 2 diabetes. Immunol Cell Biol 92:314–323Google Scholar
  13. 13.
    Creely SJ, McTernan PG, Kusminski CM et al (2007) Lipopolysaccharide activates an innate immune system response in human adipose tissue in obesity and type 2 diabetes. Am J Physiol Endocrinol Metab 292:E740–E747PubMedCrossRefGoogle Scholar
  14. 14.
    Liang H, Hussey SE, Sanchez-Avila A, Tantiwong P, Musi N (2013) Effect of lipopolysaccharide on inflammation and insulin action in human muscle. PLoS One 8:e63983PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Brun P, Castagliuolo I, Di Leo V et al (2007) Increased intestinal permeability in obese mice: new evidence in the pathogenesis of nonalcoholic steatohepatitis. Am J Physiol Gastrointest Liver Physiol 292:G518–G525PubMedCrossRefGoogle Scholar
  16. 16.
    Cucak H, Grunnet LG, Rosendahl A (2014) Accumulation of M1-like macrophages in type 2 diabetic islets is followed by a systemic shift in macrophage polarization. J Leukoc Biol 95:149–160PubMedCrossRefGoogle Scholar
  17. 17.
    Cani PD, Amar J, Iglesias MA et al (2007) Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes 56:1761–1772PubMedCrossRefGoogle Scholar
  18. 18.
    Shi H, Kokoeva MV, Inouye K, Tzameli I, Yin H, Flier JS (2006) TLR4 links innate immunity and fatty acid-induced insulin resistance. J Clin Invest 116:3015–3025PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Tsukumo DM, Carvalho-Filho MA, Carvalheira JB et al (2007) Loss-of-function mutation in Toll-like receptor 4 prevents diet-induced obesity and insulin resistance. Diabetes 56:1986–1998PubMedCrossRefGoogle Scholar
  20. 20.
    Kuo LH, Tsai PJ, Jiang MJ et al (2011) Toll-like receptor 2 deficiency improves insulin sensitivity and hepatic insulin signalling in the mouse. Diabetologia 54:168–179PubMedCrossRefGoogle Scholar
  21. 21.
    Himes RW, Smith CW (2010) Tlr2 is critical for diet-induced metabolic syndrome in a murine model. FASEB J 24:731–739Google Scholar
  22. 22.
    Poggi M, Bastelica D, Gual P et al (2007) C3H/HeJ mice carrying a toll-like receptor 4 mutation are protected against the development of insulin resistance in white adipose tissue in response to a high-fat diet. Diabetologia 50:1267–1276PubMedCrossRefGoogle Scholar
  23. 23.
    Kim F, Pham M, Luttrell I et al (2007) Toll-like receptor-4 mediates vascular inflammation and insulin resistance in diet-induced obesity. Circ Res 100:1589–1596PubMedCrossRefGoogle Scholar
  24. 24.
    Vives-Pi M, Somoza N, Fernandez-Alvarez J et al (2003) Evidence of expression of endotoxin receptors CD14, toll-like receptors TLR4 and TLR2 and associated molecule MD-2 and of sensitivity to endotoxin (LPS) in islet beta cells. Clin Exp Immunol 133:208–218PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Wen L, Peng J, Li Z, Wong FS (2004) The effect of innate immunity on autoimmune diabetes and the expression of Toll-like receptors on pancreatic islets. J Immunol 172:3173–3180PubMedCrossRefGoogle Scholar
  26. 26.
    Boni-Schnetzler M, Boller S, Debray S et al (2009) Free fatty acids induce a proinflammatory response in islets via the abundantly expressed interleukin-1 receptor I. Endocrinology 150:5218–5229PubMedCrossRefGoogle Scholar
  27. 27.
    Amyot J, Semache M, Ferdaoussi M, Fontes G, Poitout V (2012) Lipopolysaccharides impair insulin gene expression in isolated islets of Langerhans via Toll-like receptor-4 and NF-kappaB signalling. PLoS One 7:e36200PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Gibson DL, Montero M, Ropeleski MJ et al (2010) Interleukin-11 reduces TLR4-induced colitis in TLR2-deficient mice and restores intestinal STAT3 signaling. Gastroenterology 139:1277–1288PubMedCrossRefGoogle Scholar
  29. 29.
    Ii M, Matsunaga N, Hazeki K et al (2006) A novel cyclohexene derivative, ethyl (6R)-6-[N-(2-chloro-4-fluorophenyl)sulfamoyl]cyclohex-1-ene-1-carboxylate (TAK-242), selectively inhibits toll-like receptor 4-mediated cytokine production through suppression of intracellular signaling. Mol Pharmacol 69:1288–1295PubMedCrossRefGoogle Scholar
  30. 30.
    Matsunaga N, Tsuchimori N, Matsumoto T, Ii M (2011) TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol Pharmacol 79:34–41PubMedCrossRefGoogle Scholar
  31. 31.
    Takashima K, Matsunaga N, Yoshimatsu M et al (2009) Analysis of binding site for the novel small-molecule TLR4 signal transduction inhibitor TAK-242 and its therapeutic effect on mouse sepsis model. Br J Pharmacol 157:1250–1262PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    Westwell-Roper C, Dai DL, Soukhatcheva G et al (2011) IL-1 blockade attenuates islet amyloid polypeptide-induced proinflammatory cytokine release and pancreatic islet graft dysfunction. J Immunol 187:2755–2765PubMedCrossRefGoogle Scholar
  33. 33.
    Ribaux P, Ehses JA, Lin-Marq N et al (2007) Induction of CXCL1 by extracellular matrix and autocrine enhancement by IL-1 in rat pancreatic β-cells. Endocrinology 148:5582–5590PubMedCrossRefGoogle Scholar
  34. 34.
    Westwell-Roper CY, Ehses JA, Verchere CB (2014) Resident macrophages mediate islet amyloid polypeptide-induced islet IL-1beta production and beta cell dysfunction. Diabetes 63:1698–1711Google Scholar
  35. 35.
    Ellingsgaard H, Ehses JA, Hammar EB et al (2008) Interleukin-6 regulates pancreatic alpha-cell mass expansion. Proc Natl Acad Sci U S A 105:13163–13168PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Calderon B, Suri A, Miller MJ, Unanue ER (2008) Dendritic cells in islets of Langerhans constitutively present beta cell-derived peptides bound to their class II MHC molecules. Proc Natl Acad Sci U S A 105:6121–6126PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Yin N, Xu J, Ginhoux F et al (2012) Functional specialization of islet dendritic cell subsets. J Immunol 188:4921–4930PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  39. 39.
    Igoillo-Esteve M, Marselli L, Cunha DA et al (2010) Palmitate induces a pro-inflammatory response in human pancreatic islets that mimics CCL2 expression by beta cells in type 2 diabetes. Diabetologia 53:1395–1405PubMedCrossRefGoogle Scholar
  40. 40.
    Wadt KA, Larsen CM, Andersen HU, Nielsen K, Karlsen AE, Mandrup-Poulsen T (1998) Ciliary neurotrophic factor potentiates the beta-cell inhibitory effect of IL-1beta in rat pancreatic islets associated with increased nitric oxide synthesis and increased expression of inducible nitric oxide synthase. Diabetes 47:1602–1608PubMedCrossRefGoogle Scholar
  41. 41.
    Novotny GW, Lundh M, Backe MB et al (2012) Transcriptional and translational regulation of cytokine signaling in inflammatory beta-cell dysfunction and apoptosis. Arch Biochem Biophys 528:171–184PubMedCrossRefGoogle Scholar
  42. 42.
    Larsen CM, Faulenbach M, Vaag A et al (2007) Interleukin-1-receptor antagonist in type 2 diabetes mellitus. N Engl J Med 356:1517–1526PubMedCrossRefGoogle Scholar
  43. 43.
    Rissanen A, Howard CP, Botha J, Thuren T, for the Global I (2012) Effect of anti-IL-1β antibody (canakinumab) on insulin secretion rates in impaired glucose tolerance or type 2 diabetes: results of a randomized, placebo-controlled trial. Diabetes Obes Metab 14:1088–1096PubMedGoogle Scholar
  44. 44.
    Sloan-Lancaster J, Abu-Raddad E, Polzer J et al (2013) Double-blind, randomized study evaluating the glycemic and anti-inflammatory effects of subcutaneous LY2189102, a neutralizing IL-1beta antibody, in patients with type 2 diabetes. Diabetes Care 36:2239–2246PubMedCrossRefGoogle Scholar
  45. 45.
    Cavelti-Weder C, Babians-Brunner A, Keller C et al (2012) Effects of gevokizumab on glycemia and inflammatory markers in type 2 diabetes. Diabetes Care 35:1654–1662PubMedCentralPubMedCrossRefGoogle Scholar
  46. 46.
    van Asseldonk EJ, Stienstra R, Koenen TB et al (2010) The effect of the interleukin-1 cytokine family members IL-1F6 and IL-1F8 on adipocyte differentiation. Obesity 18:2234–2236PubMedCrossRefGoogle Scholar
  47. 47.
    Xoma Ltd (2011) XOMA 052 Phase 2b top line results: glucose control not demonstrated, positive anti-inflammatory effect, cardiovascular biomarker and lipid improvement and safety confirmed. Available from http://investors.xoma.com/releasedetail.cfm?ReleaseID=559470
  48. 48.
    Maedler K, Sergeev P, Ris F et al (2002) Glucose-induced beta cell production of IL-1beta contributes to glucotoxicity in human pancreatic islets. J Clin Invest 110:851–860PubMedCentralPubMedCrossRefGoogle Scholar
  49. 49.
    Masters SL, Dunne A, Subramanian SL et al (2010) Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1beta in type 2 diabetes. Nat Immunol 11:897–904PubMedCentralPubMedCrossRefGoogle Scholar
  50. 50.
    Lee HM, Kim JJ, Kim HJ, Shong M, Ku BJ, Jo EK (2013) Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes. Diabetes 62:194–204PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Dominika Nackiewicz
    • 1
  • Meixia Dan
    • 1
  • Wei He
    • 2
  • Rosa Kim
    • 1
  • Anisa Salmi
    • 1
  • Sabine Rütti
    • 3
  • Clara Westwell-Roper
    • 4
  • Amanda Cunningham
    • 1
  • Madeleine Speck
    • 1
  • Carole Schuster-Klein
    • 5
  • Beatrice Guardiola
    • 5
  • Kathrin Maedler
    • 2
  • Jan A. Ehses
    • 1
    • 6
  1. 1.Department of Surgery, Faculty of MedicineThe University of British Columbia, Child and Family Research InstituteVancouverCanada
  2. 2.Center for Biomolecular InteractionsThe University of BremenBremenGermany
  3. 3.Department of Genetic Medicine and Development, University Medical CenterUniversity of GenevaGenevaSwitzerland
  4. 4.Department of Pathology and Laboratory Medicine, Faculty of MedicineThe University of British Columbia, Child and Family Research InstituteVancouverCanada
  5. 5.ADIR – Groupe de Recherche ServierSuresnesFrance
  6. 6.Department of Cellular and Physiological Sciences, Faculty of MedicineThe University of British Columbia, Child and Family Research InstituteVancouverCanada

Personalised recommendations