Download PDF
Thromb Haemost 2014; 112(01): 151-163
DOI: 10.1160/TH13-12-1034
Endothelium and Vascular Development
Schattauer GmbH

EphrinB-mediated reverse signalling controls junctional integrity and pro-inflammatory differentiation of endothelial cells

Hui Liu
1   Institute of Physiology and Pathophysiology, Division of Cardiovascular Physiology, University of Heidelberg, Germany
,
Kavi Devraj
2   Institute of Neurology (Edinger Institute), Johann Wolfgang Goethe University Frankfurt Medical School, Frankfurt am Main, Germany
,
Kerstin Möller
1   Institute of Physiology and Pathophysiology, Division of Cardiovascular Physiology, University of Heidelberg, Germany
,
Stefan Liebner
2   Institute of Neurology (Edinger Institute), Johann Wolfgang Goethe University Frankfurt Medical School, Frankfurt am Main, Germany
,
Markus Hecker
1   Institute of Physiology and Pathophysiology, Division of Cardiovascular Physiology, University of Heidelberg, Germany
,
Thomas Korff
1   Institute of Physiology and Pathophysiology, Division of Cardiovascular Physiology, University of Heidelberg, Germany
› Author Affiliations
Further Information

Publication History

Received: 19 December 2013

Accepted after minor revision: 27 January 2014

Publication Date:
01 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

The EphB/ephrinB receptor-ligand system is pivotal for the development of the embryonic vasculature and for angiogenesis in the adult organism. We observed that (i) the expression of ephrinB2 and ephrinB1 is up-regulated in capillaries during inflammation, that (ii) these ligands are localised on the luminal endothelial surface, and that (iii) they interact with the ephrinB-receptor EphB2 on monocyte/macrophages. This study delineates the impact of ephrinB-mediated reverse signalling on the integrity and proinflammatory differentiation of the endothelium. To this end, in vitro analyses with human cultured endothelial cells reveal that knockdown of ephrinB2 or ephrinB1 impairs monocyte transmigration through the endothelium. While ephrinB2 but not ephrinB1 interacts with PECAM-1 (CD31) in this context, reverse signalling by ephrinB1 but not ephrinB2 elicits a c-Jun N-terminal kinase (JNK)-dependent up-regulation of E-selectin expression. Furthermore, treatment of endothelial cells with soluble EphB2 receptor bodies or EphB2-overexpressing mouse myeloma cells links ephrinB2 to PECAM-1 and induces its Src-dependent phosphorylation while diminishing Src homology phosphotyrosyl phosphatase-2 (SHP-2) activity and increasing endothelial cell permeability. We conclude that extravasation of EphB2 positive leukocyte populations is facilitated by lowering the integrity of endothelial cell junctions and enhancing the pro-inflammatory phenotype of the endothelium through activation of ephrinB ligands.

Keywords

Inflammation - ephrinB1 - ephrinB2 - EphB2 - monocytes - endothelial cells
 

  • References

  • 1 Schnoor M, Parkos CA. Disassembly of endothelial and epithelial junctions during leukocyte transmigration. Front Biosci 2008; 13: 6638-6652.
    CrossrefPubMedGoogle Scholar
  • 2 Engelhardt B, Wolburg H. Mini-review: Transendothelial migration of leukocytes: through the front door or around the side of the house?. Eur J Immunol 2004; 34: 2955-2963.
    CrossrefPubMedGoogle Scholar
  • 3 Augustin HG, Reiss Y. EphB receptors and ephrinB ligands: regulators of vascular assembly and homeostasis. Cell Tissue Res 2003; 314: 25-31.
    CrossrefPubMedGoogle Scholar
  • 4 Zhang J, Hughes S. Role of the ephrin and Eph receptor tyrosine kinase families in angiogenesis and development of the cardiovascular system. J Pathol 2006; 208: 453-461.
    CrossrefPubMedGoogle Scholar
  • 5 Coulthard MG, Morgan M, Woodruff TM. et al. Eph/Ephrin signaling in injury and inflammation. Am J Pathol 2012; 181: 1493-1503.
    CrossrefPubMedGoogle Scholar
  • 6 Korff T, Braun J, Pfaff D. et al. Role of ephrinB2 expression in endothelial cells during arteriogenesis: impact on smooth muscle cell migration and monocyte recruitment. Blood 2008; 112: 73-81.
    CrossrefPubMedGoogle Scholar
  • 7 Korff T, Dandekar G, Pfaff D. et al. Endothelial ephrinB2 is controlled by microenvironmental determinants and associates context-dependently with CD31. Arterioscler Thromb Vasc Biol 2006; 26: 468-474.
    CrossrefPubMedGoogle Scholar
  • 8 Yu G, Luo H, Wu Y. et al. Ephrin B2 induces T cell costimulation. J Immunol 2003; 171: 106-114.
    CrossrefPubMedGoogle Scholar
  • 9 Fuller T, Korff T, Kilian A. et al. Forward EphB4 signaling in endothelial cells controls cellular repulsion and segregation from ephrinB2 positive cells. J Cell Sci 2003; 116: 2461-2470.
    CrossrefPubMedGoogle Scholar
  • 10 Pfaff D, Heroult M, Riedel M. et al. Involvement of endothelial ephrin-B2 in adhesion and transmigration of EphB-receptor-expressing monocytes. J Cell Sci 2008; 121: 3842-3850.
    CrossrefPubMedGoogle Scholar
  • 11 Braun J, Hoffmann SC, Feldner A. et al. Endothelial cell ephrinB2-dependent activation of monocytes in arteriosclerosis. Arterioscler Thromb Vasc Biol 2011; 31: 297-305.
    CrossrefPubMedGoogle Scholar
  • 12 Kullander K, Klein R. Mechanisms and functions of Eph and ephrin signalling. Nat Rev Mol Cell Biol 2002; 03: 475-486.
    CrossrefPubMedGoogle Scholar
  • 13 Luo H, Yu G, Wu Y. et al. EphB6 crosslinking results in costimulation of T cells. J Clin Invest 2002; 110: 1141-1150.
    CrossrefPubMedGoogle Scholar
  • 14 Luo H, Yu G, Tremblay J. et al. EphB6-null mutation results in compromised T cell function. J Clin Invest 2004; 114: 1762-1773.
    CrossrefPubMedGoogle Scholar
  • 15 Luo H, Wu Z, Qi S. et al. Ephrinb1 and Ephrinb2 are associated with interleukin-7 receptor alpha and retard its internalisation from the cell surface. J Biol Chem 2011; 286: 44976-44987.
    CrossrefPubMedGoogle Scholar
  • 16 Pasquale EB. Ephephrin bidirectional signaling in physiology and disease. Cell 2008; 133: 38-52.
    CrossrefPubMedGoogle Scholar
  • 17 Rohnelt RK, Hoch G, Reiss Y. et al. Immunosurveillance modelled in vitro: naive and memory T cells spontaneously migrate across unstimulated microvascular endothelium. Int Immunol 1997; 09: 435-450.
    CrossrefPubMedGoogle Scholar
  • 18 Potente M, Gerhardt H, Carmeliet P. Basic and therapeutic aspects of angiogenesis. Cell 2011; 146: 873-887.
    CrossrefPubMedGoogle Scholar
  • 19 Muller WA. Leukocyte-endothelial-cell interactions in leukocyte transmigration and the inflammatory response. Trends Immunol 2003; 24: 327-334.
    CrossrefPubMedGoogle Scholar
  • 20 Rattan V, Sultana C, Shen Y. et al. Oxidant stress-induced transendothelial migration of monocytes is linked to phosphorylation of PECAM-1. Am J Physiol 1997; 273: E453-461.
    PubMedGoogle Scholar
  • 21 Rattan V, Shen Y, Sultana C. et al. Glucose-induced transmigration of mono-cytes is linked to phosphorylation of PECAM-1 in cultured endothelial cells. Am J Physiol 1996; 271: E711-717.
    PubMedGoogle Scholar
  • 22 Liu G, Place AT, Chen Z. et al. ICAM-1-activated Src and eNOS signaling increase endothelial cell surface PECAM-1 adhesivity and neutrophil transmigration. Blood 2012; 120: 1942-1952.
    CrossrefPubMedGoogle Scholar
  • 23 Navarro P, Ruco L, Dejana E. Differential localisation of VE- and N-cadherins in human endothelial cells: VE-cadherin competes with N-cadherin for junctional localisation. J Cell Biol 1998; 140: 1475-1484.
    CrossrefPubMedGoogle Scholar
  • 24 Stel HV, Sakariassen KS, de Groot PG. et al. Von Willebrand factor in the vessel wall mediates platelet adherence. Blood 1985; 65: 85-90.
    CrossrefPubMedGoogle Scholar
  • 25 Projahn D, Koenen RR. Platelets: key players in vascular inflammation. J Leukoc Biol 2012; 92: 1167-1175.
    CrossrefPubMedGoogle Scholar
  • 26 van Gils JM, Zwaginga JJ, Hordijk PL. Molecular and functional interactions among monocytes, platelets, and endothelial cells and their relevance for cardiovascular diseases. J Leukoc Biol 2009; 85: 195-204.
    CrossrefPubMedGoogle Scholar
  • 27 Migliaccio G, Migliaccio AR, Adamson JW. In vitro differentiation of human granulocyte/macrophage and erythroid progenitors: comparative analysis of the influence of recombinant human erythropoietin, G-CSF, GM-CSF, and IL-3 in serum-supplemented and serum-deprived cultures. Blood 1988; 72: 248-256.
    CrossrefPubMedGoogle Scholar
  • 28 Xu Z, Lai KO, Zhou HM. et al. Ephrin-B1 reverse signaling activates JNK through a novel mechanism that is independent of tyrosine phosphorylation. J Biol Chem 2003; 278: 24767-24775.
    CrossrefPubMedGoogle Scholar
  • 29 Read MA, Whitley MZ, Gupta S. et al. Tumor necrosis factor alpha-induced E-selectin expression is activated by the nuclear factor-kappaB and c-JUN N-terminal kinase/p38 mitogen-activated protein kinase pathways. J Biol Chem 1997; 272: 2753-2761.
    CrossrefPubMedGoogle Scholar
  • 30 Tai LM, Holloway KA, Male DK. et al. Amyloid-beta-induced occludin down-regulation and increased permeability in human brain endothelial cells is mediated by MAPK activation. J Cell Mol Med 2010; 14: 1101-1112.
    PubMedGoogle Scholar
  • 31 Wu JC, Yan HC, Chen WT. et al. JNK signaling pathway is required for bFGF-mediated surface cadherin downregulation on HUVEC. Exp Cell Res 2008; 314: 421-429.
    CrossrefPubMedGoogle Scholar
  • 32 Jackson JR, Seed MP, Kircher CH. et al. The codependence of angiogenesis and chronic inflammation. FASEB J 1997; 11: 457-465.
    CrossrefPubMedGoogle Scholar
  • 33 Kertesz N, Krasnoperov V, Reddy R. et al. The soluble extracellular domain of EphB4 (sEphB4) antagonizes EphB4-EphrinB2 interaction, modulates angiogenesis, and inhibits tumor growth. Blood 2006; 107: 2330-2338.
    CrossrefPubMedGoogle Scholar
  • 34 Abengozar MA, de Frutos S, Ferreiro S. et al. Blocking ephrinB2 with highly specific antibodies inhibits angiogenesis, lymphangiogenesis, and tumor growth. Blood 2012; 119: 4565-4576.
    CrossrefPubMedGoogle Scholar
  • 35 Korff T, Aufgebauer K, Hecker M. Cyclic stretch controls the expression of CD40 in endothelial cells by changing their transforming growth factor-beta1 response. Circulation 2007; 116: 2288-2297.
    CrossrefPubMedGoogle Scholar
  • 36 Ross R. Atherosclerosis--an inflammatory disease. N Engl J Med 1999; 340: 115-126.
    CrossrefPubMedGoogle Scholar
  • 37 Adams RH, Wilkinson GA, Weiss C. et al. Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev 1999; 13: 295-306.
    CrossrefPubMedGoogle Scholar
  • 38 Kojima T, Chang JH, Azar DT. Proangiogenic role of ephrinB1/EphB1 in basic fibroblast growth factor-induced corneal angiogenesis. Am J Pathol 2007; 170: 764-773.
    CrossrefPubMedGoogle Scholar
  • 39 Sakamoto A, Ishibashi-Ueda H, Sugamoto Y. et al. Expression and function of ephrin-B1 and its cognate receptor EphB2 in human atherosclerosis: from an aspect of chemotaxis. Clin Sci 2008; 114: 643-650.
    CrossrefPubMedGoogle Scholar
  • 40 Ilan N, Madri JA. PECAM-1: old friend, new partners. Curr Opin Cell Biol 2003; 15: 515-524.
    CrossrefPubMedGoogle Scholar
  • 41 Newman PJ, Newman DK. Signal transduction pathways mediated by PECAM-1: new roles for an old molecule in platelet and vascular cell biology. Arterioscler Thromb Vasc Biol 2003; 23: 953-964.
    CrossrefPubMedGoogle Scholar
  • 42 Muller WA, Weigl SA, Deng X. et al. PECAM-1 is required for transendothelial migration of leukocytes. J Exp Med 1993; 178: 449-460.
    CrossrefPubMedGoogle Scholar
  • 43 Duncan GS, Andrew DP, Takimoto H. et al. Genetic evidence for functional redundancy of Platelet/Endothelial cell adhesion molecule-1 (PECAM-1): CD31-deficient mice reveal PECAM-1-dependent and PECAM-1-independent functions. J Immunol 1999; 162: 3022-3030.
    CrossrefPubMedGoogle Scholar
  • 44 Thompson RD, Noble KE, Larbi KY. et al. Platelet-endothelial cell adhesion molecule-1 (PECAM-1)-deficient mice demonstrate a transient and cytokine-specific role for PECAM-1 in leukocyte migration through the perivascular basement membrane. Blood 2001; 97: 1854-1860.
    CrossrefPubMedGoogle Scholar
  • 45 Newman DK, Hamilton C, Newman PJ. Inhibition of antigen-receptor signaling by Platelet Endothelial Cell Adhesion Molecule-1 (CD31) requires functional ITIMs, SHP-2, and p56(lck). Blood 2001; 97: 2351-2357.
    CrossrefPubMedGoogle Scholar
  • 46 Henshall TL, Jones KL, Wilkinson R. et al. Src homology 2 domain-containing protein-tyrosine phosphatases, SHP-1 and SHP-2, are required for platelet endothelial cell adhesion molecule-1/CD31-mediated inhibitory signaling. J Immunol 2001; 166: 3098-3106.
    CrossrefPubMedGoogle Scholar
  • 47 Sagawa K, Kimura T, Swieter M. et al. The protein-tyrosine phosphatase SHP-2 associates with tyrosine-phosphorylated adhesion molecule PECAM-1 (CD31). J Biol Chem 1997; 272: 31086-31091.
    CrossrefPubMedGoogle Scholar
  • 48 Jackson DE, Ward CM, Wang R. et al. The protein-tyrosine phosphatase SHP-2 binds platelet/endothelial cell adhesion molecule-1 (PECAM-1) and forms a distinct signaling complex during platelet aggregation. Evidence for a mechanistic link between PECAM-1- and integrin-mediated cellular signaling. J Biol Chem 1997; 272: 6986-6993.
    CrossrefPubMedGoogle Scholar
  • 49 Dejana E, Orsenigo F, Lampugnani MG. The role of adherens junctions and VE-cadherin in the control of vascular permeability. J Cell Sci 2008; 121: 2115-2122.
    CrossrefPubMedGoogle Scholar
  • 50 Corada M, Liao F, Lindgren M. et al. Monoclonal antibodies directed to different regions of vascular endothelial cadherin extracellular domain affect adhesion and clustering of the protein and modulate endothelial permeability. Blood 2001; 97: 1679-1684.
    CrossrefPubMedGoogle Scholar
  • 51 Weis S, Shintani S, Weber A. et al. Src blockade stabilizes a Flk/cadherin complex, reducing edema and tissue injury following myocardial infarction. J Clin Invest 2004; 113: 885-894.
    CrossrefPubMedGoogle Scholar
  • 52 Paul R, Zhang ZG, Eliceiri BP. et al. Src deficiency or blockade of Src activity in mice provides cerebral protection following stroke. Nat Med 2001; 07: 222-227.
    CrossrefPubMedGoogle Scholar