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Thromb Haemost 1997; 77(05): 1014-1019
DOI: 10.1055/s-0038-1656095
Vessel Wall
Schattauer GmbH Stuttgart

Endothelial Cell Protein S Synthesis Is Upregulated by the Complex of IL-6 and Soluble IL-6 Receptor

W Craig Hooper
The Division of HIV/AIDS, Hematologic Diseases Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, USA
,
Donald J Phillips
The Division of HIV/AIDS, Hematologic Diseases Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, USA
,
Bruce L Evatt
The Division of HIV/AIDS, Hematologic Diseases Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, USA
› Author Affiliations
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Publication History

Received 01 August 1996

Accepted after revision 04 February 1997

Publication Date:
11 July 2018 (online)

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Summary

We have recently demonstrated that the proinflammatory cytokine, interleukin-6 (IL-6), could upregulate the production of protein S in the human hepatoma cell line, HepG-2, but not in endothelial cells. In this study, we have demonstrated that the combination of exogenous IL-6 and soluble IL-6 receptor (sIL-6R) could significantly upregulate protein S production in both primary human umbilical vein endothelial cells (HUVEC) and in the immortalized human microvascular endothelial cell line, HMEC-1. The IL-6/sIL-6R complex was also able to rapidly induce tyrosine phosphorylation of the IL-6 transducer, gpl30. Neutralizing antibodies directed against either IL-6 or gpl30 blocked protein S upregulation by the IL-6/sIL-6R complex. It was also observed that exogenous sIL-6R could also upregulate protein S by forming a complex with IL-6 constitutively produced by the endothelial cell. Two other cytokines which also utilize the gpl30 receptor, oncostatin M (OSM) and leukemia inhibitory factor (LIF), were also able to upregulate endothelial cell protein S. This study demonstrates a mechanism that allows endothelial cells to respond to IL-6 and also illustrates the potential importance of circulating soluble receptors in the regulation of the anticoagulation pathway.

 

  • References

  • 1 Heinrich PC, Castell JV, Andus T. Interleukin-6 and the acute-phase response. Biochem J 1990; 265: 621-636
    CrossrefPubMedGoogle Scholar
  • 2 Gauldie J, Geisterfer M, Richards C. Baumann H: IL-6 regulation of the hepatic acute-phase response. In: IL-6: Physiopathology and Clinical Potential. Revel M. ed. New York: Raven Press; 1992. 88. pp 153-162
    Google Scholar
  • 3 Waage A, Shalaby R, Espevik T. Tumor necrosis factor, interleukin-1, interleukin-6, interleukin-8 and interferon-gamma in septic shock. In: Cytokines in Health and Disease. Kunkel S, Remick D. (eds). New York: Marcel Dekker; 1992. pp 151-164
    Google Scholar
  • 4 Shalaby MR, Waage A, Espevik T. Cytokine regulation of interleukin 6 production by human endothelial cells. Cell Immunol 1989; 121: 372-382
    CrossrefPubMedGoogle Scholar
  • 5 Rott O, Tontsch U, Fleischer B, Cash E. Interleukin-6 production in “normal” and HTLV-1 tax expressing brain-specific endothelial cells. Eur J Immunol 1993; 23: 1987-1991
    CrossrefPubMedGoogle Scholar
  • 6 Almeida GD, Porada CD, Jeors ST, Ascensao JL. Human cytomegalovirus alters interleukin-6 production by endothelial cells. Blood 1994; 83: 370-376
    CrossrefPubMedGoogle Scholar
  • 7 Taga T, Hibi M, Hirata Y, Yamasaki K, Yasukawa K, Matsuda T, Hirano T, Kishimoto T. Interleukin-6 triggers the association of its receptor with a possible signal transducer, gpl30. Cell 1989; 58: 573-581
    CrossrefPubMedGoogle Scholar
  • 8 Hibi M, Murakami M, Saito M, Hirano T, Taga T, Kishimoto T. Molecular cloning and expression of an IL-6 signal transducer, gpl30. Cell 1990; 63: 1149-1157
    CrossrefPubMedGoogle Scholar
  • 9 Kishimoto T, Akira S, Taga T. Interleukin-6 and its receptor: A paradigm for cytokines. Science 1992; 258: 593-597
    CrossrefPubMedGoogle Scholar
  • 10 Stahl N, Yancopoulos GD. The alphas, betas and kinases of cytokine receptor complexes. Cell 1993; 74: 587-590
    CrossrefPubMedGoogle Scholar
  • 11 Gearing DP, Comeau MR, Friend DJ, Gimbel SD, Thut CJ, McGourty J, Brasher KK, King JA, Gillis S, Mosley B, Ziegler SF, Cosman D. The IL-6 signal transducer gpl30. An oncostatin M receptor and affinity converter for the LIF receptor. Science 1992; 255: 1434-1437
    CrossrefPubMedGoogle Scholar
  • 12 Zhang X-G, Gu J-J, Lu Z-Y, Yasukawa K, Yancopoulos GD, Turner K, Shoyab M, Taga T, Kishimoto T, Bataille R, Klein B. Ciliary neurotropic factor, interleukin 11, leukemia inhibitory factor, and oncostatin M are growth factors for human myeloma cell lines using the interleukin 6 signal transducer gpl30. J Exp Med 1994; 177: 1337-1342
    PubMedGoogle Scholar
  • 13 Hira Y, Taga T, Hibi M, Nakano N, Hirano T, Kishimoto T. Characterization of IL-6 receptor expression by monoclonal and polyclonal antibodies. J Immunol 1989; 143: 2900-2905
    PubMedGoogle Scholar
  • 14 Taga T, Kishimoto T. Role of a two-chain IL-6 receptor system in immune and hematopoietic cell regulation. Crit Rev Immunol 1992; 11: 265-280
    PubMedGoogle Scholar
  • 15 Mullberg J, Schooltink H, Stoyan T, Heinrich PC, Rose-John S. Protein kinase C activity is rate limiting for shedding of the interleukin-6 receptor. Biochem and Biophys Res Comm 1992; 189: 794-800
    CrossrefPubMedGoogle Scholar
  • 16 Mullberg J, Schooltink H, Stoyan T, Gunther M, Graeve L, Buse G, Mackiewicz A, Heinrich PC, Rose-John S. The soluble interleukin-6 receptor is generated by shedding. Eur J Immunol 1993; 23: 473-480
    CrossrefPubMedGoogle Scholar
  • 17 Yasukawa K, Saito T, Fukunaga T, Sekimori Y, Koishihara Y, Fukui H, Ohsugi Y, Matsuda T, Yawata H, Hirano T, Taga T, Kishimoto T. Purification and characterization of soluble human IL-6 receptor expressed in CHO cells. J. Biochem 1990; 108: 673-676
    CrossrefPubMedGoogle Scholar
  • 18 Mackiewicz A, Schooltink H, Heinrich PC, Rose-John S. Complex of soluble human IL-6-receptor/IL-6 up-regulates expression of acute-phase proteins. J Immunol 1992; 149: 2021-2027
    PubMedGoogle Scholar
  • 19 Tamura T, Udagawa N, Takahashi N, Miyaura C, Tanaka S, Yamada Y, Koishihara Y, Ohsugi Y, Kumaki K, Taga T, Kishimoto T, Suda T. Soluble interleukin-6 receptor triggers osteoclast formation by interleukin 6. Proc Nat Acad Sci USA 1993; 90: 11924-11928
    CrossrefPubMedGoogle Scholar
  • 20 van der Poll T, Biiller HR, ten Cate H, Wortel CH, Bauer KA, van DeventerSJH, Hack CE, Sauerwein HP, Rosenberg RD, ten Cate JW. Activation of coagulation after administration of tumor necrosis factor to normal subjects. New Eng J Med 1990; 322: 1622-1627
    CrossrefPubMedGoogle Scholar
  • 21 Bevilacqua MP, Prober JS, Majeau GR, Fiers W, Cotran RS, Gimbrone MA. Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the action of interleukin-1. Proc Natl Acad Sci USA 1986; 83: 4533-4537
    CrossrefPubMedGoogle Scholar
  • 22 Conway EM, Rosenberg RD. Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells. Mol Cell Biol 1988; 8: 5588-5592
    CrossrefPubMedGoogle Scholar
  • 23 Hooper WC, Phillips DJ, Ribeiro MJA, Benson JM, George VG, Ades EW, Evatt BL. Tumor Necrosis Factor-a downregulates protein S secretion in human microvascular and umbilical vein endothelial cells but not in the HepG-2 hepatoma cell line. Blood 1994; 84: 483-489
    PubMedGoogle Scholar
  • 24 Hooper WC, Phillips DJ, Evatt BL. IL-6 upregulates protein S expression in the HepG-2 hepatoma cells. Thromb Haemost 1995; 73: 819-824
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Walker FJ. Regulation of activated protein C by protein S. J Biol Chem 1981; 256: 11128-11131
    PubMedGoogle Scholar
  • 26 Taylor FB, Chang A, Esmon CT, D’Angelo A, Vigano-D’Angelo S, Blick KE. Protein C prevents the coagulopathic and lethal effect of E. coli infusion in the baboon. J Clin Invest 1987; 79: 918-925
    CrossrefPubMedGoogle Scholar
  • 27 Esmon CT, Taylor FB, Snow RT. Inflammation and coagulation: Linked processes potentially regulated through a common pathway mediated by protein C. Thromb Haemost 1991; 66: 160-165
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Taylor F, Chang FerrellG, Mather T, Catlett R, Blick K, Esmon CT. C4b-binding protein exacerbates the host response to Escherichia coli. Blood 1991; 78: 357-363
    PubMedGoogle Scholar
  • 29 Ades A, Candal FJ, Swerlick RA, George VG, Summers S, Bosse DC, Law-ley TJ. HMEC-1: Establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol 1992; 99: 683-690
    CrossrefPubMedGoogle Scholar
  • 30 Phillips DJ, Evatt BL, Hooper WC. Development of an ELISA for quantification of human protein S in cell culture fluids using commercial polyclonal antisera. J Immunoassay 1994; 15: 411-428
    PubMedGoogle Scholar
  • 31 Lai RB, Rudolph D, Buckner C, Pardi D, Hooper WC. Infection with human T-lymphotropic viruses leads to constitutive expression of leukemia inhibitory factor in interleukin-6. Blood 1993; 81: 1827-1832
    PubMedGoogle Scholar
  • 32 Podor TJ, Jirik FR, Loskutoff DJ, Carson DA. Lotz M. Human endothelial cells produce IL-6. Lack of responses to exogenous IL-6. Ann New York Acad Sci 1989; 557: 374-387
    PubMedGoogle Scholar
  • 33 Hashimoto Y, Hirohata S, Kashiwado T, Itoh K, Ishii H. Cytokine regulation of hemostatic property and IL-6 production of human endothelial cells. Inflammation 1992; 16: 613-621
    CrossrefPubMedGoogle Scholar
  • 34 Rose-John S, Schooltink H, Lenz D, Hipp E, Dufhues G, Schmitz H, Schiel X, Hirano T, Kishimoto T, Heinrich PC. Studies on the structure and regulation of the human hepatic interleukin-6 receptor. Eur J Biochem 1990; 190: 79-83
    CrossrefPubMedGoogle Scholar
  • 35 Snyers L, DeWit L, Content J. Glucocorticoid up-regulation of high-affin- ity interleukin 6 receptors on human epithelial cells. Proc Nat Acad Sci USA 1990; 87: 2838-2842
    CrossrefPubMedGoogle Scholar
  • 36 Maruo N, Morita I, Ishizaki Y, Murota S-I. Inhibitory effects of interleukin- 6 on prostaglandin 12 production in cultured bovine vascular endothelial cells. Arch Biochem and Biophys 1992; 292: 600-4
    CrossrefPubMedGoogle Scholar
  • 37 Calderon TM, Sherman J, Wilkerson H, Hatcher VB, Berman JW. Interleukin-6 modulates c-sis gene expression in cultured human endothelial cells. Cell Immunol 1992; 143: 118-126
    CrossrefPubMedGoogle Scholar
  • 38 May LT, Torcia G, Cozzolino F, Ray A, Tatter SB, Santhanam U, Sehgal PB. Interleukin-6 gene expression in human endothelial cells: RNA start sites, multiple IL-6 proteins and inhibition of proliferation. Biochem Biophys Res Comm. 1989; 159: 991-998
    PubMedGoogle Scholar
  • 39 Fair DS, Marlar RA, Levin EG. Human endothelial cells synthesize protein. S. Blood 1986; 67: 1168-1171
    PubMedGoogle Scholar
  • 40 Stern D, Brett J, Harris K, Nawroth P. Participation of endothelial cells in the protein C-protein S anticoagulant pathway: The synthesis and release of protein S. J Cell Biol 1986; 102: 1971-1978
    CrossrefPubMedGoogle Scholar
  • 41 Gaillard J-P, Bataille R, Brailly H, Zuber C, Yasukawa K, Attal M, Maruo N, Taga T, Kishimoto T, Klein B. Increased and highly stable levels of functional soluble interleukin-6 receptor in sera of patients with monoclonal gammopathy. Eur J Immunol 1993; 23: 820-824
    CrossrefPubMedGoogle Scholar
  • 42 Suzuki H, Yasukowa K, Seito T, Narazaki M, Hasegawa A, Taga T, Kishimoto T. Serum soluble interleukin-6 receptor in MRL/lpr mice is elevated with age and mediates the interleukin-6 signal. Eur J Immunol 1993; 23: 1078-1082
    CrossrefPubMedGoogle Scholar