Thromb Haemost 1992; 67(05): 578-581
DOI: 10.1055/s-0038-1648497
Original Articles
Schattauer GmbH Stuttgart

Effects of Injected Antibody against the Platelet Glycoprotein llb/llla Complex on Monkey Platelet Fibrinogen

Masahiko Suzuki
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Toshihiro Kawakatsu
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Hirokazu Nagata
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Kenjiro Hamamoto
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Koji Iwata
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Shigetoshi Ohga
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Shosaku Nomura
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Terutoshi Kokawa
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
,
Kojiro Yasunaga
The First Department of Internal Medicine, Kansai Medical University, Osaka, Japan
› Author Affiliations
Further Information

Publication History

Received 04 July 1991

Accepted after revision 03 December 1991

Publication Date:
03 July 2018 (online)

Summary

Four monkeys were injected for a 10-day period with the Fab fragment of a murine monoclonal antibody (NNKY 1-32) which inhibits the binding of fibrinogen to the platelet glycoprotein (GP) Ilb/IIIa complex. Platelet fibrinogen levels were assessed quantitatively by electroimmuno assay and qualitatively by immunoelectron microscopy.

The platelet fibrinogen level fell to 9.0 ± 2.8% of the control level after antibody administration. Immunoelectron microscopy showed that the injected antibody was localized on the inner surface of the platelet a-granule membrane.

Our findings suggest that the GP Ilb/IIIa complex can be internalized by a-granules and that it may mediate the endocytosis of plasma fibrinogen by platelets.

 
  • References

  • 1 George JN, Nurden AT, Phillips DR. Molecular defects that cause abnormalities of platelet-vessel wall interaction. N Engl J Med 1984; 311: 1084-1098
  • 2 Plow EF, McEver RP, Coller BS, Woods VL, Marguerie GA, Ginsberg MH. Related binding mechanisms for fibrinogen, fibronec-tin, von Willebrand factor and thrombospondin on thrombin-stimulated platelets. Blood 1985; 66: 724-727
  • 3 Nachman RL, Leung LLK. Complex formation of platelet membrane glycoprotein lib and/or Ilia with fibrinogen. J Clin Invest 1982; 69: 263-269
  • 4 Gogstad GO, Brosstad F, Krutanes M-B, Hagen I, Solum NO. Fibrinogen binding proteins of the human platelet glycoprotein Ilb-Illa complex: A study using crossed radio immunoelectrophoresis. Blood 1982; 60: 663-671
  • 5 McEver RP, Baenziger NL, Majerus PW. Isolation and quantitation of the platelet membrane glycoprotein deficient in thrombasthenia using a monoclonal hybridoma antibody. J Clin Invest 1980; 66: 1311-1318
  • 6 Bennett JS, Hoxie JA, Leitman SF, Vilarie G, Cines DB. Inhibition of fibrinogen binding to stimulated human platelets by a monoclonal antibody. Proc Natl Sci USA 1983; 80: 2417-2421
  • 7 Coller BS, Peerschke El, Scudder LE, Sullivan CA. A murine monoclonal antibody that completely blocks the binding of fibrinogen to platelets produces a thrombathenic-like state in normal platelets and binds to glycoprotein lib and/or Ilia. J Clin Invest 1983; 72: 325-328
  • 8 Komecki E, Tuszynski GP, Niewiarowski S. Inhibition of fibrinogen receptor-mediated platelet aggregation by heterologous anti-human platelet membrane antibody. J Biol Chem 1983; 258: 9349-9356
  • 9 McEver RP, Bennett EM, Martin MN. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein Ilb/IIIa using monoclonal antibodies. J Biol Chem 1983; 258: 5269-5275
  • 10 Pidard P, Montgomery RR, Bennett JS, Kunicki TJ. Interaction of AP-2, a monoclonal antibody specific for the human platelet glycoprotein Ilb/IIIa complex with intact platelets. J Biol Chem 1983; 258: 12582-12586
  • 11 Nomura S, Nagata H, Oda K, Kokawa T, Yusunaga K. Effects of EDTA on the membrane glycoproteins lib - Ilia complex - Analysis using flow cytometry. Thromb Res 1987; 47: 47-58
  • 12 Nomura S, Nagata H, Sone N, Oda K, Kokawa T, Yasunaga K. Analysis of platelet antigen for anti-platelet antibodies in idiopathic thrombocytopenic purpura using flow cytometry. Acta Haematol Jpn 1988; 51: 118-129
  • 13 Coller BS, Scudder LE. Inhibition of dog platelet function by in vivo infusion of F(ab’)2 fragments of a monoclonal antibody to the platelet glycoprotein Ilb/IIIa receptor. Blood 1985; 66: 1456-1459
  • 14 Coller BS, Folts JD, Scudder LE, Smith SR. Antithrombotic effects of a monoclonal antibody to the platelet glycoprotein Ilb/IIIa receptor in an experimental animal model. Blood 1986; 68: 783-786
  • 15 Hanson SR, Pareti FI, Ruggeri ZM, Marzec UM, Kunicki TJ, Montgomery RR, Zimmerman TS, Harker LA. Effects of monoclonal antibodies against the platelet glycoprotein Ilb/IIIa complex on thrombosis and hemostasis in the baboon. J Clin Invest 1988; 81: 149-158
  • 16 Hudson L, Hay FC. Cleavage of IgG by proteolytic enzymes. In: Practical Immunology. Hudson L, Hay FC. (eds) Blackwell; Oxford: 1980. p 192
  • 17 Ouchterlony O, Nilsson LA. Immunodiffusion and immunoelectrophoresis. In: Handbook of Experimental Immunology. Weir DM. (ed) Blackwell Scientific Publications; Oxford: 1978. p 19-11
  • 18 Laurell C-B. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52
  • 19 Tsutsumi H, Htay HH, Sato K, Katsumata Y. Antigenic properties of human and animal bloodstains studied by enzyme-linked immunosorbent assay (ELISA) using various antisera against specific plasma proteins. Z Rechtsmed 1987; 99: 191-196
  • 20 Ohga S, Hamamoto K, Kitajima H, Nagata H, Nomura S, Kimura T, Fujitake H, Kokawa T, Yasunaga K. The ultrastructure and distribution of fibrinogen, von Willebrand factor, and glycoprotein Ilb/IIIa complex in platelet a-granules by cryo-ultramicrotomy. Intern J Hematol 1991; 54: 331-339
  • 21 Harrison P, Wilboum B, Debili N, Vainchenker W, Breton-Gorius J, Lawrie AS, Masse J-M, Savidge GF, Cramer EM. Uptake of plasma fibrinogen into the alpha granules of human megakaryocytes and platelets. J Clin Invest 1989; 84: 1320-1324
  • 22 Handagama PJ, Shuman MA, Bainton DF. Incorporation of intravenously injected albumin, immunoglobulin G, and fibrinogen in guinea pig megakaryocyte granules. J Clin Invest 1989; 84: 73-82
  • 23 Handagama PJ, Schman MA, Bainton DF. In vivo defibrination results in markedly decreased amounts of fibrinogen in rat megakaryocytes and platelets. Am J Pathol 1990; 137: 1393-1399
  • 24 Wencel-Drake JD. Plasma membrane GP Ilb/IIIa. Evidence for a cycling receptor pool. Am J Pathol 1990; 136: 61-67
  • 25 Handagama P, Rappolee DA, Werb Z, Levin J, Bainton DF. Platelet a-granule fibrinogen, albumin, and immunoglobulin G are not synthesized by rat and mouse megakaryocytes. J Clin Invest 1990; 86: 1364-1368
  • 26 Louache F, Debili N, Cramer E, Breton-Gorius J, Vainchenker W. Fibrinogen is not synthesized by human megakaryocytes. Blood 1991; 77: 311-316
  • 27 Lange W, Luig A, Dolken G, Mertelsmann R, Kanz L. Fibrinogen ³-chain mRNA is not detected in human megakaryocytes. Blood 1991; 78: 20-25