Download PDF
Thromb Haemost 1998; 80(03): 457-462
DOI: 10.1055/s-0037-1615229
Rapid Communications
Schattauer GmbH

Differences in Platelet α-granule Release between Normals and Immune Thrombocytopenic Patients and between Young and Old Platelets

Henry M. Rinder
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Jayne B. Tracey
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Michael Recht
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Laura DeCastro
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Christine S. Rinder
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Colleen McHugh
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
,
Brian R. Smith
1   From the Departments of Laboratory Medicine, Internal Medicine (Hematology), and Pediatrics, Yale University School of Medicine, New Haven, CT, USA
› Author AffiliationsSupported by grants from the National Institutes of Health HL02668 (HMR) and HL47193 (BRS) and an American Heart Association Clinician-Scientist Award (CSR).
Further Information

Publication History

Received 01 October 1997

Accepted after resubmission 27 May 1998

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

The risk of serious bleeding in patients with immune thrombocytopenic purpura (ITP) appears to be less than in comparably thrombocytopenic patients with megakaryocytic hypoplasia. It has been proposed that this difference is due to enhanced hemostatic activity of young platelets, which are increased in the circulation during ITP. We examined α-granule release in reticulated platelets (RP), which are thought to be the youngest circulating platelets, and in older non-reticulated platelets (non-RP) in normal human controls and ITP patients. Normal controls had a mean RP of 7%, compared with 42% in ITP patients. The mean concentration of thrombin receptor agonist peptide (TRAP) causing 50% of control RP to express CD62P (EC50) was 0.82 ± 0.08 μM (SEM), significantly higher than the TRAP CD62P EC50 for RP in ITP, 0.57 ± 0.06 uM (p = 0.04). Similarly, the TRAP EC 50 for non-RP in controls, 0.84 ± 0.09 μM, was significantly higher than in ITP, 0.56 ± 0.07 μM (p = 0.03), suggesting that all platelets in ITP have an enhanced α-granule threshold response to TRAP compared with controls, while RP and older platelets within each patient group have similar threshold sensitivities to TRAP. By contrast, high-dose TRAP caused RP to express twice as much mean and total CD62P as non-RP in both ITP patients and controls (p <0.05 for both comparisons). We conclude that compared with controls, all platelets in ITP are primed to undergo α-granule release to TRAP, while in both ITP and controls, the newly circulating, reticulated platelets have the potential to contribute greater amounts of CD62P surface ligand compared with older platelets (non-RP) after stimulation. Both the increased RP% and enhanced platelet response to agonist in ITP may contribute to maintenance of hemostasis despite thrombocytopenia.

 

  • References

  • 1 Johnson CA, Abildgaard CF, Schulman I. Functional thrombocytopenia. Blood 1971; 37: 163.
    PubMedGoogle Scholar
  • 2 Karpatkin S. Heterogeneity of human platelets. II. Functional evidence suggestive of young and old platelets. J Clin Invest 1969; 48: 1083.
    CrossrefPubMedGoogle Scholar
  • 3 Shulman NR, Watkins SP, Itscoitz SB, Students AB. Evidence that the spleen retains the youngest and most hemostatically effective platelets. Trans Assoc Am Physicians 1969; 81: 302.
    PubMedGoogle Scholar
  • 4 Bussel JB, Schreiber AD. Immune thrombocytopenic purpura, neonatal alloimmune thrombocytopenia, and post-transfusion purpura. In: Hematology, Basic Principles and Practice. Hoffman R, Benz Jr. EJ, Shattil SJ, Furie B, Cohen HJ. (eds). New York: Churchill Livingstone; 1991. p 1485.
    Google Scholar
  • 5 Cieslar P, Greenberg JP, Rand ML, Packham MA, Kinlough-Rathbone RL, Mustard JF. Separation of thrombin-treated platelets from normal platelets by density gradient centrifugation. Blood 1979; 53: 867.
    PubMedGoogle Scholar
  • 6 van Oost BA, van Hien-Hagg IH, Veldhuyzen BFE, Timmermans APM, Sixma JJ. Determination of density distribution of human platelets: methodological aspects and comparison with other tests for platelet activation. Thromb Haemost 1982; 47: 239.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Ginsburg AD, Aster RH. Changes associated with platelet aging. Thromb Diath Haemorrh 1972; 27: 407.
    PubMedGoogle Scholar
  • 8 Blajchman MA, Senyi AF, Hirsh J, Genton E, George JN. Hemostatic function, survival, and membrane glycoprotein changes in young versus old rabbit platelets. J Clin Invest 1981; 68: 1289.
    CrossrefPubMedGoogle Scholar
  • 9 Hirsh J, Glynn MF, Mustard JF. The effect of platelet age on platelet adherence to collagen. J Clin Invest 1968; 47: 466.
    CrossrefPubMedGoogle Scholar
  • 10 Peng J, Friese P, Heilmann E, George JN, Burstein SA, Dale GL. Aged platelets have an impaired response to thrombin as quantitated by P-selectin expression. Blood 1994; 83: 161.
    PubMedGoogle Scholar
  • 11 Ault KA, Knowles C. In vivo biotinylation demonstrates that reticulated platelets are the youngest platelets in circulation. Exp Hematol 1995; 23: 996.
    PubMedGoogle Scholar
  • 12 Dale GL, Friese P, Hynes LA, Burstein SA. Demonstration that thiazole-orange-positive platelets in the dog are less than 24 hours old. Blood 1995; 85: 1822.
    PubMedGoogle Scholar
  • 13 Rinder HM, Munz U, Ault KA, Bonan JL, Smith BR. Utility of reticulated platelets in the evaluation of thrombopoietic disorders. Arch Pathol Lab Med 1993; 117: 606.
    PubMedGoogle Scholar
  • 14 Kienast J, Schmitz G. Flow cytometric analysis of thiazole orange uptake by platelets: a diagnostic aid in the evaluation of thrombocytopenic disorders. Blood 1990; 75: 116.
    PubMedGoogle Scholar
  • 15 Shattil SJ, Cunningham M, Hoxie JA. Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry. Blood 1987; 70: 307.
    PubMedGoogle Scholar
  • 16 Johnston GI, Pickett EB, McEver RP, George JN. Heterogeneity of platelet secretion to thrombin demonstrated by fluorescence flow cytometry. Blood 1987; 69: 1401.
    PubMedGoogle Scholar
  • 17 George JN, Woolf SH, Raskob GE, Wasser JS, Aledort LM, Ballem PJ, Blanchette VS, Bussel JB, Cines DB, Kelton JG, Lichtin AE, McMillan R, Okerbloom JA, Regan DH, Warrier I. Idiopathic thrombocytopenic purpura: a practice guideline developed by explicit methods for the American Society of Hematology. Blood 1996; 88: 3.
    PubMedGoogle Scholar
  • 18 Peterec SM, Brennan SA, Rinder HM, Wnek JL, Beardsley DS. Reticulated platelet values in normal and thrombocytopenic neonates. J Peds 1996; 129: 269.
    CrossrefPubMedGoogle Scholar
  • 19 Bonan JL, Rinder HM, Smith BR. Determination of the percentage of thiazole orange (TO)-positive “reticulated” platelets using autologous erythrocyte TO fluorescence as an internal standard. Cytometry 1993; 4: 690.
    PubMedGoogle Scholar
  • 20 Rinder HM, Bonan JL, Rinder CS, Ault KA, Smith BR. Activated and un-activated platelet adhesion to neutrophils and monocytes. Blood 1991; 78: 1760.
    PubMedGoogle Scholar
  • 21 Rinder HM, Bonan JL, Anandan S, Rinder CS, Rodrigues PA, Smith BR. Non-invasive measurement of platelet kinetics in normal and preeclamptic pregnancies. Am J Obstet Gyn 1994; 170: 117.
    CrossrefPubMedGoogle Scholar
  • 22 Joseph MA, Adams D, Maragos A, Saving KL. Flow cytometry of neonatal platelet RNA. J Ped Hematol Oncol 1996; 18: 277.
    CrossrefPubMedGoogle Scholar
  • 23 Richards EM, Baglin TP. Quantitation of reticulated platelets: methodology and clinical application. Brit J Haematol 1995; 91: 445.
    CrossrefPubMedGoogle Scholar
  • 24 Ault KA, Rinder HM, Mitchell J, Carmody MB, Vary CPH, Hillman RS. The significance of platelets with increased RNA content (reticulated platelets). Am J Clin Pathol 1992; 98: 637.
    CrossrefPubMedGoogle Scholar
  • 25 Bode AP, Orton SM, Frye MJ, Udis BJ. Vesiculation of platelets during in vitro aging. Blood 1991; 77: 887.
    PubMedGoogle Scholar
  • 26 Rinder HM, Snyder EL, Bonan JL, Napychank PA, Malkus H, Smith BR. Activation in stored platelet concentrates: correlation between membrane expression of P-selectin, glycoprotein IIb/IIIa, and β-thromboglobulin release. Transfusion 1993; 33: 25.
    CrossrefPubMedGoogle Scholar
  • 27 George JN. Platelet immunoglobulin G: its significance for the evaluation of thrombocytoepnia and for understanding the origin of α-granule proteins. Blood 1990; 76: 859.
    PubMedGoogle Scholar
  • 28 Hamburger SA, McEver RP. GMP-140 mediates adhesion of activated platelets to neutrophils. Blood 1990; 75: 550.
    PubMedGoogle Scholar
  • 29 Rinder HM, Bonan JL, Rinder CS, Ault KA, Smith BR. Dynamics of leukocyte-platelet adhesion in whole blood. Blood 1991; 78: 1730.
    PubMedGoogle Scholar
  • 30 Celi A, Pellegrini G, Lorenzet R, de Biasi A, Ready N, Furie BC, Furie B. P-selectin induces the expression of tissue factor on monocytes. Proc Natl Acad Sci USA 1994; 91: 8767.
    CrossrefPubMedGoogle Scholar
  • 31 Smith BR, Rinder HM. Interactions of platelets and endothelial cells with erythrocytes and leukocytes in thrombotic thrombocytopenic purpura. Semin Hematol 1997; 34: 90.
    PubMedGoogle Scholar
  • 32 Yanabu M, Nomura S, Fukuroi T, Suzuki M, Kawakatsu T, Kido H, Yamaguchi K, Kokawa T, Yasunaga K. Platelet activation induced by an antiplatelet autoantibody against CD9 antigen and its inhibition by another autoantibody in immune thrombocytopenic purpura. Brit J Haematol 1993; 84: 694.
    CrossrefPubMedGoogle Scholar
  • 33 Hashimoto Y, Tanabe J, Mohri H, Ohkubo T. Autoimmune antibody in a patient with idiopathic thrombocytopenic purpura reacted to the platelet low molecular weight glycoproteins and activated platelets. Am J Clin Pathol 1994; 101: 370.
    CrossrefPubMedGoogle Scholar
  • 34 Nomura S, Kido H, Yamaguchi K, Fukuroi T, Yanaba M, Kawakatsu T, Suzuki M, Kokawa T, Yasunaga K. Platelet activation by antiplatelet antibodies in immune thrombocytopenic purpura. Eur J Haematol 1992; 49: 109.
    PubMedGoogle Scholar
  • 35 Chang M, Suen Y, Meng G, Buzby JS, Bussel J, Shen V, van de Ven C, Cairo MS. Differential mechanisms in the regulation of endogenous levels of thrombopoietin and interleukin-11 during thrombocytopenia: insight into the regulation of platelet production. Blood 1996; 88: 3354.
    PubMedGoogle Scholar
  • 36 Ichikawa N, Ishida F, Shimodaira S, Tahara T, Kato T, Kitano K. Regulation of serum thrombopoietin levels by platelets and megakaryocytes in patients with aplastic anaemia and idiopathic thrombocytopenic purpura. Thromb Haemost 1996; 76: 156.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Rodriguez-Linares B, Watson SP. Thrombopoietin potentiates activation of human platelets in association with JAK2 and TYK2 phosphorylation. Biochem J 1996; 316: 93.
    CrossrefPubMedGoogle Scholar
  • 38 Wun T, Paglieroni T, Hammond WP, Kaushansky K, Foster DC. Thrombopoietin is synergistic with other hematopoietic growth factors and physiologic platelet agonists for platelet activation in vitro. Am J Hematol 1997; 54: 225.
    CrossrefPubMedGoogle Scholar
  • 39 Ballmaier M, Schulze H, Straub G, Cherkaoui K, Wittner N, Lynen S, Wolters S, Bogenberger J, Welte K. Thrombopoietin in patients with congenital thrombocytopenia and absent radii: elevated serum levels, normal receptor expression, but defective reactivity to thrombopoietin. Blood 1997; 90: 612.
    PubMedGoogle Scholar
  • 40 Oleksowicz L, Mrowiec Z, Isaacs R, Dutcher JP, Puszkin E. Morphologic and ultrastructural evidence of interleukin-6 induced platelet activation. Am J Hematol 1995; 48: 92.
    CrossrefPubMedGoogle Scholar
  • 41 Peng J, Friese P, Wolf RF, Harrison P, Downs T, Lok S, Dale GL, Burstein SA. Relative reactivity of platelets from thrombopoietin- and interleukin-6-treated dogs. Blood 1996; 87: 4158.
    PubMedGoogle Scholar