Thromb Haemost 2017; 117(02): 325-338
DOI: 10.1160/TH16-07-0553
Blood Cells, Inflammation and Infection
Schattauer GmbH

Inflammation, but not recruitment, of adipose tissue macrophages requires signalling through Mac-1 (CD11b/CD18) in diet-induced obesity (DIO)

Dennis Wolf
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
2   Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, California, USA
,
Nora Bukosza
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
David Engel
3   Department of Pathology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands
,
Marjorie Poggi
3   Department of Pathology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands
,
Felix Jehle
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
4   Atherothrombosis and Vascular Biology, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
,
Nathaly Anto Michel
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Yung-Chih Chen
4   Atherothrombosis and Vascular Biology, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
,
Christian Colberg
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Natalie Hoppe
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Bianca Dufner
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Louis Boon
5   EPIRUS Biopharmaceutical Netherlands, 3584 CM, Utrecht, The Netherlands
,
Hermann Blankenbach
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Ingo Hilgendorf
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Constantin von zur Muhlen
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Jochen Reinöhl
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Björn Sommer
6   Department of Neurosurgery, Medical Faculty of the Friedrich Alexander University of Erlangen-Nürnberg (FAU), Germany
,
Timoteo Marchini
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Mark A. Febbraio
7   Division of Diabetes & Metabolism, Garvan Institute of Medical Research, Sydney, New South Wales, Australia
,
Christian Weber
8   Institute for Cardiovascular Prevention, Ludwig Maximilians University, Munich, Germany
,
Christoph Bode
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
,
Karlheinz Peter
4   Atherothrombosis and Vascular Biology, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
,
Esther Lutgens
8   Institute for Cardiovascular Prevention, Ludwig Maximilians University, Munich, Germany
9   Department of Medical Biochemistry, Subdivision of Experimental Vascular Biology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
,
Andreas Zirlik
1   Atherogenesis Research Group, University Heart Center, University of Freiburg, Freiburg, Germany
› Author Affiliations
Further Information

Publication History

Received:20 July 2016

Accepted after major revision:18 October 2016

Publication Date:
01 December 2017 (online)

Summary

Cell accumulation is a prerequisite for adipose tissue inflammation. The leukocyte integrin Mac-1 (CD11b/CD18, αMβ2) is a classic adhesion receptor critically regulating inflammatory cell recruitment. Here, we tested the hypothesis that a genetic deficiency and a therapeutic modulation of Mac-1 regulate adipose tissue inflammation in a mouse model of diet-induced obesity (DIO). C57Bl6/J mice genetically deficient (Mac-1-/-) or competent for Mac-1 (WT) consumed a high fat diet for 20 weeks. Surprisingly, Mac-1-/- mice presented with increased diet-induced weight gain, decreased insulin sensitivity in skeletal muscle and in the liver in insulin-clamps, insulin secretion deficiency and elevated glucose levels in fasting animals, and dyslipidaemia. Unexpectedly, accumulation of adipose tissue macrophages (ATMs) was unaffected, while gene expression indicated less inflamed adipose tissue and macrophages in Mac-1-/- mice. In contrast, inflammatory gene expression at distant locations, such as in skeletal muscle, was not changed. Treatment of ATMs with an agonistic anti-Mac-1 antibody, M1/70, induced pro-inflammatory genes in cell culture. In vivo, treatment with M1/70 induced a hyper-inflammatory phenotype with increased expression of IL-6 and MCP-1, whereas accumulation of ATMs did not change. Finally, inhibition of Mac-1’s adhesive interaction to CD40L by the peptide inhibitor cM7 did not affect myeloid cell accumulation in adipose tissue. We present the surprising finding that adhesive properties of the leukocyte integrin Mac-1 are not required for macrophage accumulation in adipose tissue. Instead, Mac-1 modulates inflammatory gene expression in macrophages. These findings question the net effect of integrin blockade in cardio-metabolic disease.

D. W., N. B., and D. E. equally contributed to this work.

K. P., E. L., and A. Z. share senior authorship.

Note: The review process for this manuscript was fully handled by Gregory Y. H. Lip, Editor in Chief.

Supplementary Material to this article is available online at www.thrombosis-online.com.

 
  • References

  • 1 Moller DE, Kaufman KD. Metabolic syndrome: a clinical and molecular perspective. Ann Review Med 2005; 56: 45-62.
  • 2 Hotamisligil GS. Inflammation and metabolic disorders. Nature 2006; 444: 860-867.
  • 3 Shoelson SE, Lee J, Goldfine AB. Inflammation and insulin resistance. J Clin Invest 2006; 116: 1793-1801.
  • 4 Winer S, Winer DA. The adaptive immune system as a fundamental regulator of adipose tissue inflammation and insulin resistance. Immunol Cell Biol 2012; 90: 755-762.
  • 5 Osborn O, Olefsky JM. The cellular and signalling networks linking the immune system and metabolism in disease. Nat Med 2012; 18: 363-374.
  • 6 Weisberg SP, McCann D, Desai M. et al. Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest 2003; 112: 1796-1808.
  • 7 Lumeng CN, Bodzin JL, Saltiel AR. Obesity induces a phenotypic switch in adipose tissue macrophage polarization. J Clin Invest 2007; 117: 175-184.
  • 8 Juge-Aubry CE, Somm E, Pernin A. et al. Adipose tissue is a regulated source of interleukin-10. Cytokine 2005; 29: 270-274.
  • 9 Kanda H, Tateya S, Tamori Y. et al. MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. J Clin Invest 2006; 116: 1494-1505.
  • 10 Anderson DC, Rothlein R, Marlin SD. et al. Impaired transendothelial migration by neonatal neutrophils: abnormalities of Mac-1 (CD11b/CD18)-dependent adherence reactions. Blood 1990; 76: 2613-2621.
  • 11 Smith CW, Marlin SD, Rothlein R. et al. Cooperative interactions of LFA-1 and Mac-1 with intercellular adhesion molecule-1 in facilitating adherence and transendothelial migration of human neutrophils in vitro. J Clin Invest 1989; 83: 2008-2017.
  • 12 Flick MJ, Du X, Witte DP. et al. Leukocyte engagement of fibrin(ogen) via the integrin receptor alphaMbeta2/Mac-1 is critical for host inflammatory response in vivo. J Clin Invest 2004; 113: 1596-1606.
  • 13 Dunne JL, Ballantyne CM, Beaudet AL, Ley K. Control of leukocyte rolling velocity in TNF-alpha-induced inflammation by LFA-1 and Mac-1. Blood 2002; 99: 336-341.
  • 14 Altieri DC, Agbanyo FR, Plescia J. et al. A unique recognition site mediates the interaction of fibrinogen with the leukocyte integrin Mac-1 (CD11b/CD18). J Biol Chem 1990; 265: 12119-12122.
  • 15 Ehlers R, Ustinov V, Chen Z. et al. Targeting platelet-leukocyte interactions: identification of the integrin Mac-1 binding site for the platelet counter receptor glycoprotein Ibalpha. J Exp Med 2003; 198: 1077-1088.
  • 16 Zirlik A, Maier C, Gerdes N. et al. CD40 ligand mediates inflammation independently of CD40 by interaction with Mac-1. Circulation 2007; 115: 1571-1580.
  • 17 Wolf D, Hohmann JD, Wiedemann A. et al. Binding of CD40L to Mac-1’s I-domain involves the EQLKKSKTL motif and mediates leukocyte recruitment and atherosclerosis--but does not affect immunity and thrombosis in mice. Circ Res 2011; 109: 1269-1279.
  • 18 Shimaoka M, Springer TA. Therapeutic antagonists and conformational regulation of integrin function. Nat Rev Drug Discov 2003; 02: 703-716.
  • 19 Gonzalez-Amaro R. Cell adhesion, inflammation and therapy: old ideas and a significant step forward. Acta Pharmacol Sinica 2011; 32: 1431-1432.
  • 20 Jialal I, Adams-Huet B, Devaraj S. Monocyte cell adhesion molecule receptors in nascent metabolic syndrome. Clin Biochem 2016; 49: 505-507.
  • 21 Robker RL, Collins RG, Beaudet AL. et al. Leukocyte migration in adipose tissue of mice null for ICAM-1 and Mac-1 adhesion receptors. Obesity Res 2004; 12: 936-940.
  • 22 Dong ZM, Gutierrez-Ramos JC, Coxon A. et al. A new class of obesity genes encodes leukocyte adhesion receptors. Proc Natl Acad Sci USA 1997; 94: 7526-7530.
  • 23 Wolf D, Jehle F, Ortiz Rodriguez A. et al. CD40L deficiency attenuates diet-induced adipose tissue inflammation by impairing immune cell accumulation and production of pathogenic IgG-antibodies. PLoS One 2012; 07: e33026.
  • 24 den Boer M, Voshol PJ, Kuipers F. et al. Hepatic steatosis: a mediator of the metabolic syndrome. Lessons from animal models. Arterioscler Thromb Vasc Biol 2004; 24: 644-649.
  • 25 Sun K, Kusminski CM, Scherer PE. Adipose tissue remodelling and obesity. J Clin Invest 2011; 121: 2094-2101.
  • 26 Ley K, Laudanna C, Cybulsky MI. Nourshargh S. Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 2007; 07: 678-689.
  • 27 Winer S, Chan Y, Paltser G. et al. Normalisation of obesity-associated insulin resistance through immunotherapy. Nat Med 2009; 15: 921-929.
  • 28 Luo BH, Carman CV, Springer TA. Structural basis of integrin regulation and signaling. Annu Rev Immunol 2007; 25: 619-647.
  • 29 Ding A, Wright SD, Nathan C. Activation of mouse peritoneal macrophages by monoclonal antibodies to Mac-1 (complement receptor type 3). J Exp Med 1987; 165: 733-749.
  • 30 Simon DI. Opening the field of integrin biology to „biased agonism“. Circ Res 2011; 109: 1199-1201.
  • 31 Nishimura S, Manabe I, Nagasaki M. et al. In vivo imaging in mice reveals local cell dynamics and inflammation in obese adipose tissue. J Clin Invest 2008; 118: 710-721.
  • 32 Jiang E, Perrard XD, Yang D. et al. Essential role of CD11a in CD8+ T-cell accumulation and activation in adipose tissue. Arterioscler Thromb Vasc Biol 2014; 34: 34-43.
  • 33 Auffray C, Fogg D, Garfa M. et al. Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior. Science 2007; 317: 666-670.
  • 34 Aurora AB, Porrello ER, Tan W. et al. Macrophages are required for neonatal heart regeneration. J Clin Invest 2014; 124: 1382-1392.
  • 35 Ensan S, Li A, Besla R. et al. Self-renewing resident arterial macrophages arise from embryonic CX3CR1(+) precursors and circulating monocytes immediately after birth. Nat Immunol 2016; 17: 159-168.
  • 36 Robbins CS, Hilgendorf I, Weber GF. et al. Local proliferation dominates lesional macrophage accumulation in atherosclerosis. Nat Med 2013; 19: 1166-1172.
  • 37 Drechsler M, Megens RT, van Zandvoort M. et al. Hyperlipidaemia-triggered neutrophilia promotes early atherosclerosis. Circulation 2010; 122: 1837-1845.
  • 38 Soehnlein O. Multiple roles for neutrophils in atherosclerosis. Circ Res 2012; 110: 875-888.
  • 39 Yakubenko VP, Bhattacharjee A, Pluskota E, Cathcart MK. alphaMbeta(2) integrin activation prevents alternative activation of human and murine macrophages and impedes foam cell formation. Circ Res 2011; 108: 544-554.
  • 40 Maiguel D, Faridi MH, Wei C. et al. Small molecule-mediated activation of the integrin CD11b/CD18 reduces inflammatory disease. Science Signal 2011; 04: ra57.
  • 41 Kubo N, Boisvert WA, Ballantyne CM, Curtiss LK. Leukocyte CD11b expression is not essential for the development of atherosclerosis in mice. J Lipid Res 2000; 41: 1060-1066.
  • 42 Talukdar S, Oh da Y, Bandyopadhyay G. et al. Neutrophils mediate insulin resistance in mice fed a high-fat diet through secreted elastase. Nat Med 2012; 18: 1407-1412.
  • 43 Jakubzick C, Gautier EL, Gibbings SL. et al. Minimal differentiation of classical monocytes as they survey steady-state tissues and transport antigen to lymph nodes. Immunity 2013; 39: 599-610.
  • 44 Nishimura S, Manabe I, Nagasaki M. et al. CD8+ effector T cells contribute to macrophage recruitment and adipose tissue inflammation in obesity. Nat Med 2009; 15: 914-920.
  • 45 Poggi M, Engel D, Christ A. et al. CD40L deficiency ameliorates adipose tissue inflammation and metabolic manifestations of obesity in mice. Arterioscler Thromb Vasc Biol 2011; 31: 2251-2260.
  • 46 Kim CS, Kim JG, Lee BJ. et al. Deficiency for costimulatory receptor 4-1BB protects against obesity-induced inflammation and metabolic disorders. Diabetes 2011; 60: 3159-3168.
  • 47 Chatzigeorgiou A, Seijkens T, Zarzycka B. et al. Blocking CD40-TRAF6 signalling is a therapeutic target in obesity-associated insulin resistance. Proc Natl Acad Sci USA 2014; 111: 2686-2691.
  • 48 Wolf D, Jehle F, Anto Michel N. et al. Co-Inhibitory Suppression of T Cell Activation by CD40 Protects from Obesity and Adipose Tissue Inflammation in Mice. Circulation 2014; 129: 2414-2425.
  • 49 Huang W, Metlakunta A, Dedousis N. et al. Depletion of liver Kupffer cells prevents the development of diet-induced hepatic steatosis and insulin resistance. Diabetes 2010; 59: 347-357.
  • 50 Tessem JS, Jensen JN, Pelli H. et al. Critical roles for macrophages in islet angiogenesis and maintenance during pancreatic degeneration. Diabetes 2008; 57: 1605-1617.
  • 51 Lech M, Anders HJ. Macrophages and fibrosis: How resident and infiltrating mononuclear phagocytes orchestrate all phases of tissue injury and repair. Biochim Biophys Acta 2013; 1832: 989-997.
  • 52 Cao C, Lawrence DA, Strickland DK, Zhang L. A specific role of integrin Mac-1 in accelerated macrophage efflux to the lymphatics. Blood 2005; 106: 3234-3241.
  • 53 Johannsen DL, Tchoukalova Y, Tam CS. et al. Effect of 8 weeks of overfeeding on ectopic fat deposition and insulin sensitivity: testing the „adipose tissue expandability“ hypothesis. Diabetes Care 2014; 37: 2789-2797.
  • 54 Wernstedt Asterholm I, Tao C, Morley TS. et al. Adipocyte inflammation is essential for healthy adipose tissue expansion and remodelling. Cell Metabol 2014; 20: 103-118.
  • 55 Mauer J, Chaurasia B, Goldau J. et al. Signalling by IL-6 promotes alternative activation of macrophages to limit endotoxemia and obesity-associated resistance to insulin. Nat Immunol 2014; 15: 423-430.