PDF herunterladen
Thromb Haemost 1995; 74(02): 764-769
DOI: 10.1055/s-0038-1649810
Original Article
Vessel Wall
Schattauer GmbH Stuttgart

Characterization of Endothelial Cell Differential Attachment to Fibrin and Fibrinogen and Its Inhibition by Arg-Gly-Asp-Containing Peptides

Mei-Chi Chang
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
,
Bih-Ru Wang
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
,
Tur-Fu Huang
The Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 25. August 1994

Accepted 30. März 1995

Publikationsdatum:
06. Juli 2018 (online)

Auch verfügbar auf
  als PDF herunterladen  Lizenzen und Reprints

Summary

We investigated the adhesion of human umbilical vein endothelial cells (HUVECs) to fibrin(ogen) molecule of varying structure for identifying sites that mediate cell attachment. Fibrin was prepared either with ancrod which liberates only FPA from fibrinogen, or with thrombin, which liberates both FPA and FPB. Both fibrin preparations equally supported HUVEC attachment. GRGDS, RGD-containing peptides of snake venoms, and monoclonal antibodies against αvβ3 (23C6 and 7E3) inhibited the attachment of HUVECs to fibrin by 65–75%. In contrast, the attachment of HUVECs to fibrinogen was less effective and was almost completely inhibited by both RGD-containing peptides and by antibodies against integrin αvβ3 (85-95% inhibition). The C-terminal dodecapeptide of fibrinogen γ chain (residues 400–411) inhibited minimally the attachment of HUVECs to fibrin. Additionally, the binding of RGD-containing snake venom peptides to HUVECs was both RGD- and divalent-cation-dependent. The IC50s for inhibition of HUVEC attachment to fibrin were 0.09 μM (rhodostomin), 1.54 μM (trigramin) and 1.64 μM (halysin).

These results indicate that fibrin mediated support of cell attachment is independent of the cleavage of FPB from fibrinogen. HUVEC attachment to fibrinogen was almost completely inhibited by RGD-containing peptides and by antibodies against αvβ3. In contrast, the attachment to fibrin was partially resistant to RGD-containing peptides and to the monoclonal antibodies against integrin αvβ3. However, αvβ3 is the major receptor mediating HUVEC attachment to fibrin.

 

  • References

  • 1 Clark RAF, Dvorak HF, Clovin RB. Fibronectin in delayed-type hypersensitivity skin reactions: associations with vessel permeability and endothelial cell activation. J Immunol 1981; 126: 787-793
    PubMedSuche in Google Scholar
  • 2 Clark RAF, Della PelleP, Manseau E, Lanigan JM, Dvorak HF, Clovin RB. Blood vessel fibronectin increases in conjunction with endothelial cell proliferation and capillary ingrowth during wound healing. J Invest Dermatol 1982; 79: 269-276
    CrossrefPubMedSuche in Google Scholar
  • 3 Tanaka K, Sueishi K. Biology of disease: the coagulation and fibrinolysis systems and atherosclerosis. Lab Invest 1993; 69: 5-18
    PubMedSuche in Google Scholar
  • 4 Olander JV, Bremer ME, Marasa JC, Feder J. Fibrin-enhanced endothelial cell organization. J Cell Physiol 1985; 125: 1-9
    CrossrefPubMedSuche in Google Scholar
  • 5 Watanabe K, Tanaka K. Influence of fibrin, fibrinogen and fibrinogen degradation products on cultured endothelial cells. Atherosclerosis 1983; 44: 57-70
    PubMedSuche in Google Scholar
  • 6 Weimar B, Delvos U. The mechanism of fibrin-induced disorganization of cultured human endothelial cell monolayers. Arteriosclerosis 1986; 6: 139-145
    CrossrefPubMedSuche in Google Scholar
  • 7 Ribes JA, Francis CW, Wagner DD. Fibrin induces release of von Wille- brand factor from endothelial cells. J Clin Invest 1987; 79: 117-123
    CrossrefPubMedSuche in Google Scholar
  • 8 Bunce LA, Sporn LA, Francis CW. Endothelial cell spreading on fibrin requires fibrinopeptide B cleavage and amino acid residues 15-42 of the β chain. J Clin Invest 1992; 89: 842-850
    CrossrefPubMedSuche in Google Scholar
  • 9 Francis CW, Bunce LA, Sporn LA. Endothelial cell responses to fibrin mediated by FPB cleavage and the amino terminus of the β chain. Blood Cells 1993; 19: 291-307
    PubMedSuche in Google Scholar
  • 10 Hynes RO. Integrins: a family of cell surface receptors. Cell 1987; 48: 549-554
    CrossrefPubMedSuche in Google Scholar
  • 11 Dejana E. Endothelial cell adhesive receptors. J Cardiovasc Pharmacol 1993; 21 Suppl (Suppl. 01) S18-S21
    PubMedSuche in Google Scholar
  • 12 Charo IF, Bekeart LS, Phillips DR. Platelet glycoprotein IIb-IIIa-like proteins mediate endothelial cell attachment to adhesive proteins and the extracellular matrix. J Biol Chem 1987; 262: 9935-9938
    PubMedSuche in Google Scholar
  • 13 Teng CM, Huang TF. Inventory of exogenous inhibitors of platelet aggregation. Thromb Haemost 1991; 65: 624-626
    Thieme ConnectPubMedSuche in Google Scholar
  • 14 Huang TF, Ouyang C, Teng CM. Rhodostomin, a snake venom peptide and its fragment inhibit platelet aggregation by acting as fibrinogen receptor antagonist. 11th International Congress on Thrombosis, Ljubljana, Yugoslavia 1990; abstract 141
    PubMedSuche in Google Scholar
  • 15 Huang TF, Holt JC, Lukasiewicz H, Niewiarowski S. Trigramin, a low molecular weight peptide inhibiting fibrinogen interaction with platelet receptors expressed on glycoprotein IIb/IIIa complex. J Biol Chem 1987; 262: 16157-16163
    PubMedSuche in Google Scholar
  • 16 Huang TF, Liu CZ, Ouyang C, Teng CM. Halysin, an antiplatelet Arg-Gly-Asp-containing snake venom peptide, as fibrinogen receptor antagonist. Biochem Pharmacol 1991; 42: 1209-1219
    CrossrefPubMedSuche in Google Scholar
  • 17 Charo IF, Bekeart LS, Phillips DR. Platelet glycoprotein IIb-IIIa-like proteins mediate endothelial cell attachment to adhesive proteins and the extracellular matrix. J Biol Chem 1987; 262: 9935-9938
    PubMedSuche in Google Scholar
  • 18 Grossi IM, Hatfield JS, Fitzgerald LA, Newcome M, Taylor JD, Honn KV. Role of tumor cell glycoproteins immunologically related to glycoprotein lb and IIb/IIIb in tumor cell-platelet and tumor cell-matrix interactions. FASEBJ 1988; 2: 2385-2395
    CrossrefPubMedSuche in Google Scholar
  • 19 Martinez J, Rich E, Barsigian C. Transglutaminase-mediated cross-linking of fibrinogen by human umbilical vein endothelial cells. J Biol Chem 1989; 264: 20502-20508
    PubMedSuche in Google Scholar
  • 20 Chang MC, Huang TF. In vivo effect of a thrombin-like enzyme on platelet plug formation induced in mesenteric microvessels of mice. Thromb Res 1994; 73: 31-38
    CrossrefPubMedSuche in Google Scholar
  • 21 Au LC, Lin SB, Chou JS, Teh GW, Chang KJ, Shih CC. Molecular cloning and sequence analysis of thecDNA for ancrod, a thrombin-like enzyme from the venom of Calloselasma rhodostoma. Biochem J 1993; 294: 387-390
    CrossrefPubMedSuche in Google Scholar
  • 22 Johnson JF, Seegers WH. Techniques in blood coagulation used at Wayne University. 1953; 42-44
    PubMedSuche in Google Scholar
  • 23 Jaffe EA, Nachman RL, Beeker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedSuche in Google Scholar
  • 24 Tapparelli C, Metternich R, Ehrhardt C, Cook NS. Synthetic low-molecular weight thrombin inhibitors: molecular design and pharmacological profile. TiPS 1993; 14: 366-376
    PubMedSuche in Google Scholar
  • 25 Sprandio D, Shapiro SS, Thiagarajan P, McCord S. Cultured human umbilical vein endothelial cells contain a membrane glycoprotein iinnumologically related to platelet glycoprotein Ib. Blood 1988; 71: 234-237
    PubMedSuche in Google Scholar
  • 26 Asch AS, Adelman B, Fujimoto M, Nachman RL. Identification and isolation of a platelet GP Ib-like protein in human umbilical vein endothelial cells and bovine aortic smooth muscle cells. J Clin Invest 1988; 81: 1600-1607
    CrossrefPubMedSuche in Google Scholar
  • 27 Tranqui L, Andrieux A, Hudry-Clergeon G, Ryekewaert JJ, Serger S, Chapel A, Ginsberg MH, Plow EF, Marguerie G. Differential structural requirements for fibrinogen binding to platelets and to endothelial cells. J Cell Biol 1989; 108: 2519-2527
    CrossrefPubMedSuche in Google Scholar
  • 28 Smith JW, Ruggeri ZM, Kunick TJ, Cheresh DA. Interaction of integrins αvβ3and glycoprotein IIb-IIIa with fibrinogen. J Biol Chem 1990; 265: 12267-12271
    PubMedSuche in Google Scholar
  • 29 Cheresh DA, Berliner SA, Vicente V, Ruggeri ZM. Recognition of distinct adhesive sites on fibrinogen by related integrins on platelets and endothelial cells. Cell 1989; 58: 945-953
    CrossrefPubMedSuche in Google Scholar
  • 30 Ewart MR, Hatton MW C, Basford JM, Dodgson KS. The proteolytic action of Arvin on human fibrinogen. Biochem J 1970; 118: 603-609
    CrossrefPubMedSuche in Google Scholar
  • 31 Pizzo SV, Schwartz WL, Hill RL, McKee PA. Mechanism of anerod anticoagulation. J Clin Invest 1972; 51: 2841-2849
    CrossrefPubMedSuche in Google Scholar
  • 32 Doolittle RF. Fibrinogen and fibrin. Annu Rev Biochem 1984; 53: 195-229
    CrossrefPubMedSuche in Google Scholar
  • 33 Hamaguchi M, Bunee LA, Sporn LA, Francis CW. Spreading of platelets on fibrin is mediated by the amino terminus of the β chain including peptides β15-42. Blood 1993; 81: 2348-2356
    PubMedSuche in Google Scholar
  • 34 Chiu TH, Nyials E, Tureotte LR. Microealorimetric and electrophoretic studies of protein sorption from plasma. Trans Am Soe Aerif Int Organs 1978; 24: 389-403
    PubMedSuche in Google Scholar
  • 35 Tomikawa M, Iwamoto M, Olsson P, Soderman S, Blomhack B. On the platelet-fibrinogen interaction. Thromb Res 1980; 19: 869-876
    CrossrefPubMedSuche in Google Scholar
  • 36 Lindon JN, McManama G, Kushner L, Merrill EW, Salzman EW. Does the conformation of adsorbed fibrinogen dictate platelet interactions with artilical surfaces. Blood 1986; 68: 355-362
    PubMedSuche in Google Scholar
  • 37 Lawler J, Weinstein R, Hynes RO. Cell attachment to thrombospondin: the role of Arg-Gly-Asp, calcium, and integrin receptors. J Cell Biol 1988; 107: 2351-2361
    CrossrefPubMedSuche in Google Scholar