PDF herunterladen
Thromb Haemost 1992; 68(05): 600-605
DOI: 10.1055/s-0038-1646324
Original Article
Schattauer GmbH Stuttgart

Quantitation of Platelet Glycoprotein IV (CD36) in Healthy Subjects and in Patients with Essential Thrombocythemia Using an Immunocapture Assay

Valérie Thibert
1   The INSERM U 353, Hôpital St. Louis, Paris, France
,
Sylvia Bellucci
2   The Département d'Angio-Hématologie, Hôpital Lariboisière, Paris, France
,
Lena Edelman
3   The Institut Pasteur, Paris, France
,
Narendra N Tandon
4   The American Red Cross, Cell Biology Laboratory, Rockville, MD, USA
,
Chantal Legrand
1   The INSERM U 353, Hôpital St. Louis, Paris, France
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 05. März 1992

Accepted after revision 22. Juni 1992

Publikationsdatum:
04. Juli 2018 (online)

Auch verfügbar auf
  als PDF herunterladen  Lizenzen und Reprints

Summary

Glycoprotein IV (GPIIIb, CD36) is a major platelet membrane glycoprotein which is thought to participate in a number of adhesive reactions and to mediate signal transduction. In order to measure the total content of GPIV in human platelets, we have developed a simple and sensitive solid-phase radioimmunoassay based on the immunocapture of GPIV from Triton X-100-solubilized platelets. FA6-152, a monoclonal antibody to GPIV was coated on microtiter plates and bound antigen was quantified with a radiolabeled polyclonal antibody to GPIV Using purified GPIV as a standard, the coefficients of variation of the assay were found to be less than 10% at concentrations of GPIV ranging from 0.15 to 0.75 µg/ml. The assay was validated by the parallelism obtained between purified GPIV dose-response curves and those obtained with platelet lysates, indicating a similar antigenic activity for GPIV in both samples. The level of GPIV in platelets from healthy donors was 0.23 ± 0.05 (mean ± SD, n = 15) µg per 100 µg of platelet proteins and a mean value of 27,440 ± 6,200 (SD) molecules per platelet was calculated. The radioimmunoassay could be used to discriminate between the high level of platelet GPIV in patients with essential thrombocythemia (mean ± SD = 81,850 ± 27,780 molecules/platelet; n = 8) and the normal GPIV level in patients with secondary thrombocytosis (mean ± SD = 26,810 ± 4,030 molecules/platelet; n = 5), thereby demonstrating the clinical usefulness of the assay. The specific increase in platelet GPIV in patients with essential thrombocythemia was confirmed by immunoblot analysis whereas no increase in platelet GPIb or GPIIb-IIIa was observed by this technique.

 

  • REFERENCES

  • 1 Clemetson KJ, Mc Gregor JL. Characterization of platelet membrane glycoproteins. In: Platelets in Biology and Pathology III. MacIntyre DE, Gordon JL. (eds) Elsevier Science Publishers BV, Amsterdam: 1987. pp 160-212
    Suche in Google Scholar
  • 2 Barnwell JW, Ockenhouse CF, Knowles DM. Monoclonal antibody OKM5 inhibits the in vitro binding of Plasmodium falciparum-infected erythrocytes to monocytes, endothelial, and C32 melanoma cells. J Immunol. 1985; 135: 3494-3497
    PubMedSuche in Google Scholar
  • 3 Edelman P, Vinci G, Villeval JL, Vainchenker W, Henri A, Mig-lierina R, Rouger P, Reviron P, Breton-Gorius J, Sureau C, Edelman L. A monoclonal antibody against an erythrocyte ontogenic antigen identifies fetal and adult erythroid progenitors. Blood 1986; 67: 56-63
    PubMedSuche in Google Scholar
  • 4 Kieffer N, Bettaieb A, Legrand C, Coulombel L, Vainchenker W, Edelman L, Breton-Gorius J. Developmentally regulated expression of a 78 kDa erythroblast membrane glycoprotein immunologically related to the platelet thrombospondin receptor. Biochem J 1989; 262: 835-842
    CrossrefPubMedSuche in Google Scholar
  • 5 Greenwalt DE, Watt KWK, So OY, Jiwani N. PAS IV, an integral membrane protein of mammary epithelial cells, is related to platelet and endothelial cell CD36 (GPIV). Biochemistry 1990; 29: 7054-7059
    CrossrefPubMedSuche in Google Scholar
  • 6 Asch AS, Barnwell J, Silverstein RL, Nachman RL. Isolation of the thrombospondin membrane receptor. J Clin Invest 1987; 79: 1054-1061
    CrossrefPubMedSuche in Google Scholar
  • 7 McGregor JL, Catimel B, Parmentier S, Clezardin P, Dechavanne M, Leung LLK. Rapid purification and partial characterization of human platelet glycoprotein IIIb. J Biol Chem 1989; 264: 501-506
    PubMedSuche in Google Scholar
  • 8 Tandon NN, Lipsky RH, Burgess WH, Jamieson GA. Isolation and characterization of human platelet GPIV (CD36). J Biol Chem 1989; 264: 7570-7575
    PubMedSuche in Google Scholar
  • 9 Oquendo P, Hundt E, Lawler J, Seed B. CD36 directly mediates cytoadherence of Plasmodium falciparum parasitized erythrocytes. Cell. 1989; 58: 095-101
    PubMedSuche in Google Scholar
  • 10 Tandon NN, Kralisz U, Jamieson GA. Identification of glycoprotein IV (CD36) as a primary receptor for platelet-collagen adhesion. J Biol Chem 1989; 264: 7576-7583
    PubMedSuche in Google Scholar
  • 11 Kieffer N, Nurden AT, Hasitz M, Titeux M, Breton-Gorius J. Identification of platelet membrane thrombospondin binding molecules using an anti-thrombospondin antibody. Biochim Biophys Acta 1988; 967: 408-415
    CrossrefPubMedSuche in Google Scholar
  • 12 Beiso P, Pidard D, Fournier D, Dubernard V, Legrand C. Studies on the interaction of platelet glycoprotein IIb-IIIa and glycoprotein IV with fibrinogen and thrombospondin: A new immunological approach. Biochim Biophys Acta 1990; 1033: 07-12
    CrossrefPubMedSuche in Google Scholar
  • 13 Leung LLK. Role of thrombospondin in platelet aggregation. J Clin Invest 1984; 74: 1764-1772
    CrossrefPubMedSuche in Google Scholar
  • 14 Tuszynski GP, Rothman VL, Murphy A, Siegler K, Knudsen KA. Thrombospondin promotes platelet aggregation. Blood 1988; 72: 109-115
    PubMedSuche in Google Scholar
  • 15 Yamamoto N, Ikeda H, Tandon NN, Herman J, Tomiyama Y, Mitani T, Sekiguchi S, Lipsky R, Kralisz U, Jamieson GA. A platelet membrane glycoprotein (GP) deficiency in healthy blood donors: Naka-platelets lack detectable GPIV (CD36). Blood. 1990; 76: 1698-1703
    PubMedSuche in Google Scholar
  • 16 Kehrel B, Kronenberg A, Schwippert B, Niesing-Bresch D, Niehues U, Tschöpe D, van de Loo J, Clemetson KJ. Thrombospondin binds normally to glycoprotein IIIb deficient platelets. Biochem Biophys Res Commun 1991; 179: 985-991
    CrossrefPubMedSuche in Google Scholar
  • 17 Tandon NN, Ockenhouse CF, Greco NJ, Jamieson GA. Adhesive functions of platelets lacking glycoprotein IV (CD36). Blood 1991; 78: 2809-2813
    PubMedSuche in Google Scholar
  • 18 Ockenhouse CF, Magowan C, Chulay JD. Activation of monocytes and platelets by monoclonal antibodies or infected erythrocytes binding to the CD36 surface receptor in vitro. J Clin Invest 1989; 84: 468-475
    CrossrefPubMedSuche in Google Scholar
  • 19 Aiken ML, Ginsberg MH, Byers-Ward V, Plow EF. Effects of OKM5, a monoclonal antibody to glycoprotein IV, on platelet aggregation and thrombospondin surface expression. Blood 1990; 76: 2501-2509
    PubMedSuche in Google Scholar
  • 20 Legrand C, Pidard D, Beiso P, Tenza D, Edelman L. Interaction of a monoclonal antibody to glycoprotein IV (CD36) with human platelets and its effect on platelet function. Platelets 1991; 2: 099-105
    CrossrefPubMedSuche in Google Scholar
  • 21 Huang MM, Bolen JB, Barnwell JW, Shattil SJ, Brugge JS. Membrane glycoprotein IV (CD36) is physically associated with the Fyn, Lyn, and Yes protein-tyrosine kinases in human platelets. Proc Natl Acad Sci USA 1991; 88: 7844-7848
    CrossrefPubMedSuche in Google Scholar
  • 22 Bolin RB, Okumura T, Jamieson GA. Changes in distribution of platelet membrane glycoproteins in patients with myeloproliferative disorders. Am J Hematol 1977; 3: 63-71
    CrossrefPubMedSuche in Google Scholar
  • 23 Eche N, Sie P, Caranobe C, Nouvel J, Pris J, Boneu B. Platelets in myeloproliferative disorders. III: Glycoprotein profile in relation to platelet function and platelet density. Scand J Haematol 1981; 26: 123-129
    PubMedSuche in Google Scholar
  • 24 Clezardin P, Mc Gregor JL, Dechavanne M, Clemetson KJ. Platelet membrane glycoprotein abnormalities in patients with myeloprolifera tive disorders and secondary thrombocytosis. Br J Haematol 1985; 60: 331-344
    CrossrefPubMedSuche in Google Scholar
  • 25 Legrand C, Bellucci S, Disdier M, Edelman L, Tobelem G. Platelet thrombospondin and glycoprotein IV abnormalities in patients with essential thrombocythemia. Effect of α-interferon treatment. Am J Hematol 1991; 38: 307-313
    CrossrefPubMedSuche in Google Scholar
  • 26 Bellucci S, Janvier M, Tobelem G, Flandrin G, Charpak Y, Berger R, Boiron M. Essential thrombocythemias. Clinical evolutionary and biological data. Cancer 1986; 58: 2440-2447
    CrossrefPubMedSuche in Google Scholar
  • 27 Hunter WM, Greenwood FC. Preparation of iodine 131 labeled human growth hormone of high specific activity. Nature 1962; 194: 495-496
    CrossrefPubMedSuche in Google Scholar
  • 28 Wicki AN, Clemetson KJ. Structure and function of platelet membrane glycoprotein Ib and V. Effects of leukocyte elastase and other proteases on platelets response to von Willebrand factor and thrombin. Eur J Biochem 1985; 153: 01-11
    CrossrefPubMedSuche in Google Scholar
  • 29 Cramer EM, Savidge GF, Vainchenker W, Berndt MC, Pidard D, Caen JP, Massé JM, Breton-Gorius J. Alpha-granule pool of glycoprotein IIb-IIIa in normal and pathologic platelets and megakaryocytes. Blood 1990; 75: 1220-1227
    PubMedSuche in Google Scholar
  • 30 Legrand C, Dubernard V, Caen J. Platelet aggregation: its relation with ADP-induced fibrinogen binding to platelets and ADP-related membrane enzyme activities. Eur J Biochem 1984; 142: 465-471
    CrossrefPubMedSuche in Google Scholar
  • 31 Markwell MAK, Haas SM, Lieber LL, Tolbert NE. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 1978; 87: 206-210
    CrossrefPubMedSuche in Google Scholar
  • 32 Laemmli UK. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedSuche in Google Scholar
  • 33 Kieffer N, Boizard B, Didry D, Wautier JL, Nurden AT. Immunochemical characterization of the platelet-specific alloantigen Leka. A comparative study with the PLA1 alloantigen. Blood. 1984; 64: 1212-1219
    PubMedSuche in Google Scholar
  • 34 Legrand C, Thibert V, Dubernard V, Bégault B, Lawler J. Molecular requirements for the interaction of thrombospondin with thrombin-activated human platelets. Modulation of platelet aggregation. Blood 1992; 79: 1995-2003
    PubMedSuche in Google Scholar
  • 35 Yamazaki H, Suzuki H, Yamamoto N, Baba I, Arai M, Fujimaki M, Tanoue K. Glycoprotein (GP) IV localizes on plasma membrane including open canalicular system but not alpha-granule membrane. Thromb Haemostas 1991; 65: 1070a
    PubMedSuche in Google Scholar