Download PDF
Thromb Haemost 1998; 79(06): 1106-1110
DOI: 10.1055/s-0037-1615024
Rapid Communication
Schattauer GmbH

Evaluation of Thrombopoiesis in Thrombocytopenic Disorders by Simultaneous Measurement of Reticulated Platelets of Whole Blood and Serum Thrombopoietin Concentrations

Yukako Koike
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Akiko Yoneyama
2   Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo, Tokyo, Japan
,
Jun Shirai
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Tateru Ishida
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Eriko Shoda
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Kouji Miyazaki
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Sinji Sunaga
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Ryoichi Horie
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Katunori Aoki
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Kazuhiko Koike
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Iturou Ogata
1   From the First Department of Internal Medicine, Tokyo, Japan
,
Tomoyuki Tahara
4   Pharmaceutical Reseach Laboratory, Kirin Brewery Co. Ltd., Maebashi, Japan
,
Takashi Kato
4   Pharmaceutical Reseach Laboratory, Kirin Brewery Co. Ltd., Maebashi, Japan
,
Kazuhiko Nakahara
2   Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo, Tokyo, Japan
,
Toshitugu Kariya
3   Division of Haematology, Tokyo Kosei Nenkin Hospital, Tokyo, Japan
,
Masaaki Higashihara
5   Fourth Department of Internal Medicine, Kitasato University, School of Medicine, Kanagawa, Japan
› Author Affiliations
Further Information

Publication History

Received 27 May 1997

Accepted after resubmission 06 February 1998

Publication Date:
07 December 2017 (online)

    Permissions and Reprints

Summary

To evaluate thrombopoiesis in thrombocytopenic disorders, we simultaneously determined reticulated platelet counts in whole blood by FACScan flow cytometry and serum thrombopoietin (TPO) concentrations by a sensitive sandwich ELISA. The subjects were 40 healthy volunteers and 45 thrombocytopenic patients. In idiopathic thrombocytopenic purpura (ITP), the percentage of reticulated platelets was significantly elevated (5.61 ± 2.02%: mean ± SD) relative to normal controls (2.17 ± 0.90%), but serum TPO concentrations (1.91 ± 1.27 fmol/l) did not differ significantly from the normal range (1.43 ± 0.62 fmol/l). The patients with aplastic anemia (AA) had decreased reticulated platelet counts and markedly increased serum TPO concentrations (13.65 ± 10.64 fmol/l). In thrombocytopenic patients with liver cirrhosis (LC), the absolute number of reticulated platelets (1.65 ± 1.11 × 109/l) decreased similarly that in AA. However, serum TPO concentrations (1.38 ± 0.50 fmol/l) did not increase in contrast to AA. Our findings suggested a possible dual mechanism of thrombocytopenia in LC; that is, thrombocytopenia in LC results from the decreased TPO production primarily in the liver adding to an increase in platelet sequestration in the spleen.

 

  • References

  • 1 Ingram M, Coopersmith A. Reticulated platelets following acute blood loss.. Br J Haematol 1969; 17: 225-9.
    CrossrefPubMedGoogle Scholar
  • 2 Lee LG, Chen CH, Chiu LA. Thiazole orange: a new dye for reticulocyte analysis.. Cytometry 1986; 7: 508-17.
    CrossrefPubMedGoogle Scholar
  • 3 Newman PJ, Gorski J, White GC, Gidwitz S, Cretney CJ, Aster RH. Enzymatic amplification of platelet-specific messenger RNA using the polymerase chain reaction.. J Clin Invest 1988; 82: 739-43.
    CrossrefPubMedGoogle Scholar
  • 4 Sottile J, Mosher DF, Fullenweider J, George JN. Human platelets contain mRNA transcripts for platelet factor 4 and actin.. Thromb Haemost 1989; 62: 1100-2.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 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: 116-21.
    PubMedGoogle Scholar
  • 6 Ault KA, Rinder HM, Mitchell J, Carmody MB, Vary CPH, Hillman RS. The significance of platelets with increased RNA content (reticulated platelets): a measure of the rate of thrombopoiesis.. Am J Clin Pathol 1992; 98: 637-46.
    CrossrefPubMedGoogle Scholar
  • 7 de Sauvage FJ, Hass PE, Spencer SD, Malloy BE, Gurney AL, Spencer SA, Darbonne WC, Henzel WJ, Wong SC, Kuang WJ, Oles KJ, Hultgren B, Solberg Jr LA, Goeddel DV, Eaton DL. Stimulation of megakaryocytopoiesis and thrombopoiesis by the c-Mpl ligand.. Nature 1994; 369: 533-8.
    CrossrefPubMedGoogle Scholar
  • 8 Bartley TD, Bogenberger J, Hunt P, Li YS, Lu HS, Martin F, Chang MS, Samal B, Nichol JL, Swift S, Johnson MJ, Hsu RY, Parker VP, Suggs S, Skrine JD, Merewether LA, Clogston C, Hsu E, Hokom MM, Hornkohl A, Choi E, Pangelinan M, Sun Y, Mar V, McNinch J, Simonet L, Jacobsen F, Xie C, Shutter J, Chute H, Basu R, Selander L, Trollinger D, Sieu L, Padilla D, Trail G, Elliott G, Izumi R, Covey T, Crouse J, Garcia A, Xu W, Del Castillo J, Biron J, Cole S, Hu MCT, Pacifici R, Ponting I, Saris C, Wen D, Yung YP, Lin H, Bosselman RA. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl.. Cell 1994; 77: 1117-24.
    CrossrefPubMedGoogle Scholar
  • 9 Sohma Y, Akahori H, Seki N, Hori T, Ogami K, Kato T, Shimada Y, Kawamura K, Miyazaki H. Molecular cloning and chromosomal localization of the human thrombopoietin gene.. FEBS Lett 1994; 353: 57-61.
    CrossrefPubMedGoogle Scholar
  • 10 Kuter DJ, Beeler DL, Rosenberg RD. The purification of megapoietin: a physiological regulator of megakaryocyte growth and platelet production.. Proc Natl Acad Sci U S A 1994; 91: 11104-8.
    CrossrefPubMedGoogle Scholar
  • 11 Lok S, Kaushansky K, Holly RD, Kuijper JL, Lofton-Day CE, Oort PJ, Grant FJ, Heipel MD, Burkhead SK, Kramer JM, Bell LA, Sprecher CA, Blumberg H, Johnson R, Prunkard D, Ching AFT, Mathewes SL, Bailey MC, Forstrom JW, Buddle MM, Osborn SG, Evans SJ, Sheppard PO, Presnell SR, O’Hara PJ, Hagen FS, Roth GJ, Foster DC. Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo.. Nature 1994; 369: 565-8.
    CrossrefPubMedGoogle Scholar
  • 12 Kaushansky K, Lok S, Holly RD, Broudy VC, Lin N, Bailey MC, Forstrom JW, Buddle MM, Oort PJ, Hagen FS, Roth GJ, Papayannopoulou T, Foster DC. Promotion of megakaryocyte progenitor expansion and differentiation by the c-Mpl ligand thrombopoietin.. Nature 1994; 369: 568-71.
    CrossrefPubMedGoogle Scholar
  • 13 Wendling F, Maraskovsky E, Debili N, Florindo C, Teepe M, Titeux M, Methia N, Breton-Gorius J, Cosman D, Vainchenker W. c-Mpl ligand is a humoral regulator of megakaryocytopoiesis.. Nature 1994; 369: 571-4.
    CrossrefPubMedGoogle Scholar
  • 14 Shimada Y, Kato T, Ogami K, Horie K, Kokubo A, Kudo Y, Maeda E, Sohma Y, Akahori H, Kawamura K, Miyazaki H. Production of thrombopoietin (TPO) by rat hepatocytes and hepatoma cell lines.. Exp Hematol 1995; 23: 1388-96.
    PubMedGoogle Scholar
  • 15 Tahara T, Usuki K, Sato H, Ohashi H, Morita H, Tsumura H, Matsumoto A, Miyazaki H, Urabe A, Kato T. A sensitive sandwich ELISA for measuring thrombopoietin in human serum: serum thrombopoietin levels in healthy volunteers and in patients with haemopoietic disorders.. Br J Haematol 1996; 93: 783-8.
    CrossrefPubMedGoogle Scholar
  • 16 Tassies D, Reverter JC, Cases A, Escolar G, Villamor N, Lopez-Pedret J, Castillo R, Ordinas A. Reticulated platelets in uremic patients: effect of hemodialysis and continuous ambulatory peritoneal dialysis.. Am J Hematol 1995; 50: 161-6.
    CrossrefPubMedGoogle Scholar
  • 17 Watanabe K, Takeuchi K, Kawai Y, Ikeda Y, Kubota F, Nakamoto H. Automated measurement of reticulated platelets in estimating thrombopoiesis.. Eur J Haematol 1995; 54: 163-71.
    PubMedGoogle Scholar
  • 18 Kuter DJ, Rosenberg RD. The reciprocal relationship of thrombopoietin (c-Mpl ligand) to changes in the platelet mass during busulfan-induced thrombocytopenia in the rabbit.. Blood 1995; 85: 2720-30.
    PubMedGoogle Scholar
  • 19 Emmons RVB, Reid DM, Cohen RL, Meng G, Young NS, Dunbar CE, Shulman NR. Human thrombopoietin levels are high when thrombocytopenia is due to megakaryocyte deficiency and low when due to increased platelet destruction.. Blood 1996; 87: 4068-71.
    PubMedGoogle Scholar
  • 20 Aster RH. Pooling of platelets in the spleen: role in the pathogenesis of “hypersplenic” thrombocytopenia.. J Clin Invest 1966; 45: 645-57.
    CrossrefPubMedGoogle Scholar
  • 21 Wright CS, Doan CA, Bouroncle BA, Zollinger RM. Direct splenic arterial and venous blood studies in the hypersplenic syndromes before and after epinephrine.. Blood 1951; 6: 195-212.
    PubMedGoogle Scholar
  • 22 Aoki Y, Hirai K, Tanikawa K. Mechanism of thrombocytopenia in liver cirrhosis: kinetics of indium-111 tropolone labelled platelets.. Eur J Nucl Med 1993; 20: 123-9.
    PubMedGoogle Scholar
  • 23 Noguchi H, Hirai K, Aoki Y, Satake K, Tanikawa K. Changes in platelet kinetics after a partial splenic arterial embolization in cirrhotic patients with hypersplenism.. Hepatology 1995; 22: 1682-8.
    CrossrefPubMedGoogle Scholar
  • 24 Dameshek W. Hypersplenism.. Bull New York Acad Med 1955; 31: 113-36.
    PubMedGoogle Scholar
  • 25 Cohen P, Gardner FH, Barnett GO. Reclassification of the thrombocytopenias by the Cr51-labeling method for measuring platelet life span.. N Engl J Med 1961; 264: 1294-9.
    CrossrefPubMedGoogle Scholar
  • 26 Mayer M, Herrmann I, Kempgens U, Queisser W. In vivo labeling of platelets with 75Se-selenomethionine in patients with hepatic cirrhosis and thrombocytopenia.. Thromb Haemost 1977; 37: 47-52.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Ohashi T, Fujiwara K. Symposium on blood disorder in systemic diseases (5) hematologic abnormalities in liver disease.. (Japanese) Nippon Naika Gakkai Zasshi 1982; 71: 1677-81.
    CrossrefPubMedGoogle Scholar
  • 28 Shimodaira S, Ishida F, Ichikawa N, Tahara T, Kato T, Kodaira H, Ito T, Tanaka E, Sodeyama T, Kiyosawa K, Kitano K. Serum thrombopoietin (c-Mpl ligand) levels in patients with liver cirrhosis.. Thromb Haemost 1996; 76: 545-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Ulich TR, del Castillo J, Yin S, Swift S, Padilla D, Senaldi G, Bennett L, Shutter J, Bogenberger J, Sun D, Samal B, Shimamoto G, Lee R, Steinbrink R, Boone T, Sheridan WT, Hunt P. Megakaryocyte growth and development factor ameliorates carboplatin-induced thrombocytopenia in mice.. Blood 1995; 86: 971-6.
    PubMedGoogle Scholar
  • 30 Stoffel R, Wiestner A, Skoda RC. Thrombopoietin in thrombocytopenic mice: evidence against regulation at the mRNA level and for a direct regulatory role of platelets.. Blood 1996; 87: 567-73.
    PubMedGoogle Scholar
  • 31 McCarty JM, Sprugel KH, Fox NE, Sabath DE, Kaushansky K. Murine thrombopoietin mRNA levels are modulated by platelet count.. Blood 1995; 86: 3668-75.
    PubMedGoogle Scholar
  • 32 Fielder PJ, Gurney AL, Stefanich E, Marian M, Moore MW, Carver-Moore K, de Sauvage FJ. Regulation of thrombopoietin levels by c-mpl-mediated binding to platelets.. Blood 1996; 87: 2154-61.
    PubMedGoogle Scholar