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MIF is a noncognate ligand of CXC chemokine receptors in inflammatory and atherogenic cell recruitment

Nature Medicine volume 13, pages 587–596 (2007)Cite this article

Abstract

The cytokine macrophage migration inhibitory factor (MIF) plays a critical role in inflammatory diseases and atherogenesis. We identify the chemokine receptors CXCR2 and CXCR4 as functional receptors for MIF. MIF triggered Gαi- and integrin-dependent arrest and chemotaxis of monocytes and T cells, rapid integrin activation and calcium influx through CXCR2 or CXCR4. MIF competed with cognate ligands for CXCR4 and CXCR2 binding, and directly bound to CXCR2. CXCR2 and CD74 formed a receptor complex, and monocyte arrest elicited by MIF in inflamed or atherosclerotic arteries involved both CXCR2 and CD74. In vivo, Mif deficiency impaired monocyte adhesion to the arterial wall in atherosclerosis-prone mice, and MIF-induced leukocyte recruitment required Il8rb (which encodes Cxcr2). Blockade of Mif but not of canonical ligands of Cxcr2 or Cxcr4 in mice with advanced atherosclerosis led to plaque regression and reduced monocyte and T-cell content in plaques. By activating both CXCR2 and CXCR4, MIF displays chemokine-like functions and acts as a major regulator of inflammatory cell recruitment and atherogenesis. Targeting MIF in individuals with manifest atherosclerosis can potentially be used to treat this condition.

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Figure 1: MIF-triggered mononuclear cell arrest is mediated by CXCR2, CXCR4 and CD74.
Figure 2: MIF-triggered mononuclear cell chemotaxis is mediated by CXCR2, CXCR4 and CD74.
Figure 3: MIF triggers rapid integrin activation and calcium signaling.
Figure 4: MIF interaction with CXCR2 or CXCR4 and formation of a CXCR2-CD74 complex.
Figure 5: MIF-driven monocyte arrest in inflamed or atherosclerotic arteries involves CXCR2.
Figure 6: MIF-induced atherogenic and microvascular inflammation through CXCR2 in vivo and effects of MIF blockade on plaque regression.

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References

  1. Baggiolini, M., Dewald, B. & Moser, B. Interleukin-8 and related chemotactic cytokines–CXC and CC chemokines. Adv. Immunol. 55, 97–179 (1994).

    Article  CAS  Google Scholar 

  2. Charo, I.F. & Ransohoff, R.M. The many roles of chemokines and chemokine receptors in inflammation. N. Engl. J. Med. 354, 610–621 (2006).

    Article  CAS  Google Scholar 

  3. Murphy, P.M. et al. International union of pharmacology. XXII. Nomenclature for chemokine receptors. Pharmacol. Rev. 52, 145–176 (2000).

    CAS  PubMed  Google Scholar 

  4. Rot, A. & von Andrian, U.H. Chemokines in innate and adaptive host defense: basic chemokinese grammar for immune cells. Annu. Rev. Immunol. 22, 891–928 (2004).

    Article  CAS  Google Scholar 

  5. Weber, C., Schober, A. & Zernecke, A. Chemokines: key regulators of mononuclear cell recruitment in atherosclerotic vascular disease. Arterioscler. Thromb. Vasc. Biol. 24, 1997–2008 (2004).

    Article  CAS  Google Scholar 

  6. Libby, P. Inflammation in atherosclerosis. Nature 420, 868–874 (2002).

    Article  CAS  Google Scholar 

  7. Hansson, G.K. Inflammation, atherosclerosis, and coronary artery disease. N. Engl. J. Med. 352, 1685–1695 (2005).

    Article  CAS  Google Scholar 

  8. Tedgui, A. & Mallat, Z. Cytokines in atherosclerosis: pathogenic and regulatory pathways. Physiol. Rev. 86, 515–581 (2006).

    Article  CAS  Google Scholar 

  9. Laudanna, C. & Alon, R. Right on the spot. Chemokine triggering of integrin-mediated arrest of rolling leukocytes. Thromb. Haemost. 95, 5–11 (2006).

    Article  CAS  Google Scholar 

  10. Degryse, B. & deVirgilio, M. The nuclear protein HMGB1, a new kind of chemokine? FEBS Lett. 553, 11–17 (2003).

    Article  CAS  Google Scholar 

  11. Yang, D. et al. β-defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science 286, 525–528 (1999).

    Article  CAS  Google Scholar 

  12. Sun, H.W., Bernhagen, J., Bucala, R. & Lolis, E. Crystal structure at 2.6 Å resolution of human macrophage migration inhibitory factor. Proc. Natl. Acad. Sci. USA 93, 5191–5196 (1996).

    Article  CAS  Google Scholar 

  13. Donnelly, S.C. et al. Regulatory role for macrophage migration inhibitory factor in acute respiratory distress syndrome. Nat. Med. 3, 320–323 (1997).

    Article  CAS  Google Scholar 

  14. Morand, E.F., Leech, M. & Bernhagen, J. MIF: a new cytokine link between rheumatoid arthritis and atherosclerosis. Nat. Rev. Drug Discov. 5, 399–410 (2006).

    Article  CAS  Google Scholar 

  15. Bernhagen, J. et al. MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia. Nature 365, 756–759 (1993).

    Article  CAS  Google Scholar 

  16. Calandra, T. & Roger, T. Macrophage migration inhibitory factor: a regulator of innate immunity. Nat. Rev. Immunol. 3, 791–800 (2003).

    Article  CAS  Google Scholar 

  17. Kleemann, R. et al. Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1. Nature 408, 211–216 (2000).

    Article  CAS  Google Scholar 

  18. Leng, L. et al. MIF signal transduction initiated by binding to CD74. J. Exp. Med. 197, 1467–1476 (2003).

    Article  CAS  Google Scholar 

  19. Shi, X. et al. CD44 is the signaling component of the macrophage migration inhibitory factor-CD74 receptor complex. Immunity 25, 595–606 (2006).

    Article  CAS  Google Scholar 

  20. David, J.R. Delayed hypersensitivity in vitro: its mediation by cell-free substances formed by lymphoid cell-antigen interaction. Proc. Natl. Acad. Sci. USA 56, 72–77 (1966).

    Article  CAS  Google Scholar 

  21. Lan, H.Y. et al. The pathogenic role of macrophage migration inhibitory factor in immunologically induced kidney disease in the rat. J. Exp. Med. 185, 1455–1465 (1997).

    Article  CAS  Google Scholar 

  22. Gregory, J.L. et al. Reduced leukocyte-endothelial cell interactions in the inflamed microcirculation of macrophage migration inhibitory factor-deficient mice. Arthritis Rheum. 50, 3023–3034 (2004).

    Article  CAS  Google Scholar 

  23. Schrans-Stassen, B.H.G.J., Lue, H., Sonnemans, D.G.P., Bernhagen, J. & Post, M.J. Stimulation of vascular smooth muscle cell migration by macrophage migration inhibitory factor (MIF). Antioxid. Redox Signal. 7, 1211–1216 (2005).

    Article  CAS  Google Scholar 

  24. Burger-Kentischer, A. et al. Expression of macrophage migration inhibitory factor in different stages of human atherosclerosis. Circulation 105, 1561–1566 (2002).

    Article  CAS  Google Scholar 

  25. Lin, S.G. et al. De novo expression of macrophage migration inhibitory factor in atherogenesis in rabbits. Circ. Res. 87, 1202–1208 (2000).

    Article  CAS  Google Scholar 

  26. Schober, A. et al. Stabilization of atherosclerotic plaques by blockade of macrophage migration inhibitory factor after vascular injury in apolipoprotein E-deficient mice. Circulation 109, 380–385 (2004).

    Article  CAS  Google Scholar 

  27. Kong, Y.Z. et al. Macrophage migration inhibitory factor induces MMP-9 expression: implications for destabilization of human atherosclerotic plaques. Atherosclerosis 178, 207–215 (2005).

    Article  CAS  Google Scholar 

  28. Pan, J.H. et al. Macrophage migration inhibitory factor deficiency impairs atherosclerosis in low-density lipoprotein receptor-deficient mice. Circulation 109, 3149–3153 (2004).

    Article  CAS  Google Scholar 

  29. Huo, Y. et al. The chemokine KC, but not monocyte chemoattractant protein-1, triggers monocyte arrest on early atherosclerotic endothelium. J. Clin. Invest. 108, 1307–1314 (2001).

    Article  CAS  Google Scholar 

  30. Weber, K.S., von Hundelshausen, P., Clark-Lewis, I., Weber, P.C. & Weber, C. Differential immobilization and hierarchical involvement of chemokines in monocyte arrest and transmigration on inflamed endothelium in shear flow. Eur. J. Immunol. 29, 700–712 (1999).

    Article  CAS  Google Scholar 

  31. Shamri, R. et al. Lymphocyte arrest requires instantaneous induction of an extended LFA-1 conformation mediated by endothelium-bound chemokines. Nat. Immunol. 6, 497–506 (2005).

    Article  CAS  Google Scholar 

  32. Jones, S.A., Dewald, B., Clark-Lewis, I. & Baggiolini, M. Chemokine antagonists that discriminate between interleukin-8 receptors. Selective blockers of CXCR2. J. Biol. Chem. 272, 16166–16169 (1997).

    Article  CAS  Google Scholar 

  33. Bertini, R. et al. Thioredoxin, a redox enzyme released in infection and inflammation, is a unique chemoattractant for neutrophils, monocytes, and T cells. J. Exp. Med. 189, 1783–1789 (1999).

    Article  CAS  Google Scholar 

  34. Wakasugi, K. & Schimmel, P. Two distinct cytokines released from a human aminoacyl-tRNA synthetase. Science 284, 147–151 (1999).

    Article  CAS  Google Scholar 

  35. Weber, C. & Koenen, R.R. Fine-tuning leukocyte responses: towards a chemokine 'interactome'. Trends Immunol. 27, 268–273 (2006).

    Article  CAS  Google Scholar 

  36. Waters, C.M., Connell, M.C., Pyne, S. & Pyne, N.J. c-Src is involved in regulating signal transmission from PDGFβ receptor-GPCR(s) complexes in mammalian cells. Cell. Signal. 17, 263–277 (2005).

    Article  CAS  Google Scholar 

  37. Roscic-Mrkic, B. et al. RANTES (CCL5) uses the proteoglycan CD44 as an auxiliary receptor to mediate cellular activation signals and HIV-1 enhancement. Blood 102, 1169–1177 (2003).

    Article  CAS  Google Scholar 

  38. Boisvert, W.A. et al. Up-regulated expression of the CXCR2 ligand KC/GRO-α in atherosclerotic lesions plays a central role in macrophage accumulation and lesion progression. Am. J. Pathol. 168, 1385–1395 (2006).

    Article  CAS  Google Scholar 

  39. Navab, M. et al. Human apolipoprotein A-I and A-I mimetic peptides: potential for atherosclerosis reversal. Curr. Opin. Lipidol. 15, 645–649 (2004).

    Article  CAS  Google Scholar 

  40. Proudfoot, A.E. Chemokine receptors: multifaceted therapeutic targets. Nat. Rev. Immunol. 2, 106–115 (2002).

    Article  CAS  Google Scholar 

  41. Weber, C., Aepfelbacher, M., Haag, H., Ziegler-Heitbrock, H.W. & Weber, P.C. TNF induces enhanced responses to platelet-activating factor and differentiation in human monocytic Mono Mac 6 cells. Eur. J. Immunol. 23, 852–859 (1993).

    Article  CAS  Google Scholar 

  42. Ostermann, G., Weber, K.S., Zernecke, A., Schroder, A. & Weber, C. JAM-1 is a ligand of the β(2) integrin LFA-1 involved in transendothelial migration of leukocytes. Nat. Immunol. 3, 151–158 (2002).

    Article  CAS  Google Scholar 

  43. Ben-Baruch, A. et al. IL-8 and NAP-2 differ in their capacities to bind and chemoattract 293 cells transfected with either IL-8 receptor type A or type B. Cytokine 9, 37–45 (1997).

    Article  CAS  Google Scholar 

  44. Weber, C., Alon, R., Moser, B. & Springer, T.A. Sequential regulation of α4β1 and α5β1 integrin avidity by CC chemokines in monocytes: implications for transendothelial chemotaxis. J. Cell Biol. 134, 1063–1073 (1996).

    Article  CAS  Google Scholar 

  45. Kleemann, R., Grell, M., Mischke, R., Zimmermann, G. & Bernhagen, J. Receptor-binding and cellular uptake studies of macrophage migration inhibitory factor (MIF): use of biologically active MIF derivatives. J. Interferon Cytokine Res. 22, 351–363 (2002).

    Article  CAS  Google Scholar 

  46. Hayashi, S. et al. Synthetic hexa- and heptapeptides that inhibit IL-8 from binding to and activating human blood neutrophils. J. Immunol. 154, 814–824 (1995).

    CAS  PubMed  Google Scholar 

  47. Fingerle-Rowson, G. et al. The p53-dependent effects of macrophage migration inhibitory factor revealed by gene targeting. Proc. Natl. Acad. Sci. USA 100, 9354–9359 (2003).

    Article  CAS  Google Scholar 

  48. Verschuren, L. et al. Effect of low dose atorvastatin versus diet-induced cholesterol lowering on atherosclerotic lesion progression and inflammation in apolipoprotein E*3-Leiden transgenic mice. Arterioscler. Thromb. Vasc. Biol. 25, 161–167 (2005).

    Article  CAS  Google Scholar 

  49. Keane, M.P., Belperio, J.A., Xue, Y.Y., Burdick, M.D. & Strieter, R.M. Depletion of CXCR2 inhibits tumor growth and angiogenesis in a murine model of lung cancer. J. Immunol. 172, 2853–2860 (2004).

    Article  CAS  Google Scholar 

  50. Zernecke, A. et al. SDF-1α/CXCR4 axis is instrumental in neointimal hyperplasia and recruitment of smooth muscle progenitor cells. Circ. Res. 96, 784–791 (2005).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank E. Liehn, S. Knarren and L. Verschuren for assistance with the atherosclerotic mouse models; S. Kraemer for help with internalization assays; A. Ben-Baruch (Department of Cell Research and Immunology, Tel Aviv University) for HEK293-CXCR2 cells; H.W.L. Ziegler-Heitbrock (University of Leicester) for MonoMac6 cells; M. Locati (Istituto Clinico Humanitas) for L1.2 cells; H. Hengel (University of Düsseldorf) for SVECs; A. Ludwig and E. Brandt (Department of Immunology and Cell Biology, Forschungszentrum Borstel) for CXCR2 antibody RII115; D. Staunton (Department of Biomedical Engineering, Genome and Biomedical Sciences Facility, University of California at Davis) for the antibody to 327C; Anormed Inc. (Genzyme Corporate Offices) for AMD3465; and A. Ludwig, E. Morand, M. Thelen and A. Kapurniotu for helpful discussions. Supported by the Deutsche Forschungsgemeinschaft grants BE 1977/2-1, BE 1977/4-1 (J.B. and C.W.), WE 1913/7-1 (C.W.), SFB542-A7 (J.B.) and SFB542-C12 (C.W.); US National Institutes of Health grants AI43210 and AR49610 (R.B.); Australian National Health and Medical Research Council (NHMRC) program grant 334067 (M.J.H. and S.R.M.); US NIH grant AR51807-01 (M.J.H. and S.R.M.); and the Netherlands Organization for Scientific Research grant VENI 016.036.061 (R.K.).

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Author notes

  1. Jürgen Bernhagen and Christian Weber: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Cell Biology, Institute of Biochemistry, University Hospital Aachen, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, D-52074, Germany

    Jürgen Bernhagen, Hongqi Lue, Manfred Dewor & Ivan Georgiev

  2. Institute of Molecular Cardiovascular Research, University Hospital Aachen, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, D-52074, Germany

    Regina Krohn, Alma Zernecke, Rory R Koenen & Christian Weber

  3. Centre for Inflammatory Diseases, Monash University, Victoria, 3168, Australia

    Julia L Gregory & Michael J Hickey

  4. Medical Policlinic, Ludwig-Maximilians-University, Munich, D-80336, Germany

    Andreas Schober

  5. Department of Medicine, Pathology, Epidemiology and Public Health, The Anlyan Center, Yale University School of Medicine, New Haven, 06520, Connecticut, USA

    Lin Leng & Richard Bucala

  6. Department of Vascular and Metabolic Diseases, TNO Quality of Life, Gaubius Laboratory/Biosciences, Leiden, 2301 CE, The Netherlands

    Teake Kooistra & Robert Kleemann

  7. Department of Hematology and Oncology, Medical Clinic I, University Hospital Cologne, Cologne, D-50937, Germany

    Günter Fingerle-Rowson

  8. Mario Negri-Institute for Pharmacological Research, Milan, 20157, Italy

    Pietro Ghezzi

  9. Department of Vascular Surgery, Leiden University Medical Center, Leiden, 2333 CK, The Netherlands

    Robert Kleemann

  10. School of Molecular and Biomedical Science, University of Adelaide, Adelaide, 5005, SA, Australia

    Shaun R McColl

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Correspondence to Jürgen Bernhagen or Christian Weber.

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Competing interests

J.B., R.B., and L.L. are co-inventors on patents describing the use of MIF inhibition for the treatment of inflammatory disease.

Supplementary information

Supplementary Fig. 1

Structural homology between the CXCL8 dimer and the MIF monomer. (PDF 72 kb)

Supplementary Fig. 2

Adhesion and chemotaxis assays with various cellular and antibody controls. (PDF 52 kb)

Supplementary Fig. 3

Ex vivo perfusion of murine carotid arteries. (PDF 122 kb)

Supplementary Fig. 4

The monoclonal Mif antibody NIHIII.D.9 specifically recognizes Mif, but not Cxcl1/Kc or CXCL8. (PDF 109 kb)

Supplementary Video 1

This movie shows an ex vivo perfusion of the carotid artery of an Apoe−/− mouse fed an atherogenic diet for 6 weeks with calcein-AM-labeled MonoMac6 cells. Applying stroboscopic epifluorescence illumination, monocytic cells firmly adherent to the vessel wall, rolling along the artery wall and flowing at the rate of perfusion (4 μL/min) can be visualized in different segments of the perfused artery. The movie is a representative of a control artery corresponding to the data in Fig. 5a-f and Supplementary Fig. 3. (MPG 2854 kb)

Supplementary Video 2

MIF-mediated accumulation of leukocytes in carotid arteries of chimeric mice in vivo relies on CXCR2 as revealed by intravital microscopy. The first sequence in this video shows several leukocytes labeled by intravenous injection of rhodamine, which have become adherent to the carotid artery wall 4 h after intraperitoneal injection of Tnf-α in a wild-type Mif+/+ mouse reconstituted with wild-type bone marrow (Mif+/+/WT bone marrow). The following sequences show very few adherent leukocytes in the carotid arteries of wild-type mice reconstituted with Cxcr2−/−/ BM (Mif+/+/Cxcr2−/− bone marrow) or Mif−/− mice reconstituted with wild-type BM (Mif−/−/WT bone marrow) and no further reduction was seen in Mif−/− mice reconstituted with Cxcr2−/− BM (Mif−/−/Cxcr2−/− bone marrow). The movie corresponds to the data in Fig. 5g,h. (MOV 997 kb)

Supplementary Video 3

MIF induces CXCR2-dependent leukocyte recruitment in vivo in the microvasculature of the cremaster muscle visualized by intravital microscopy. The first sequence in this video shows a cremasteric postcapillary venule after a control (saline) injection (saline control). Some leukocytes are visible rolling within the venule, but no leukocytes are adherent within the venule and very few leukocytes are apparent in the extravascular tissue. The second sequence in this video shows a postcapillary venule after local injection of MIF (1μ g) and IgG isotype control antibody (MIF + control IgG). While leukocyte rolling and blood flow are not markedly altered compared with saline-injected mice, many leukocytes are visible adherent to the endothelial surface. Several leukocytes have been recruited to the tissue and are visible adjacent to the postcapillary venule. The last sequence in this video shows the effects of an antibody to Cxcr2 on MIF (1 μg)-induced leukocyte adhesion and emigration (MIF + anti-Cxcr2). The numbers of adherent leukocytes within the venule are dramatically reduced compared with mice treated with MIF alone and very few leukocytes have been recruited to the extravascular tissue. The movie corresponds to the data in Fig. 6b,c. (MOV 1468 kb)

Supplementary Methods (PDF 165 kb)

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Bernhagen, J., Krohn, R., Lue, H. et al. MIF is a noncognate ligand of CXC chemokine receptors in inflammatory and atherogenic cell recruitment. Nat Med 13, 587–596 (2007). https://doi.org/10.1038/nm1567

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