Download PDF
Thromb Haemost 1995; 73(03): 506-513
DOI: 10.1055/s-0038-1653805
Original Articles
Platelets
Schattauer GmbH Stuttgart

Anticoagulant-induced Pseudothrombocytopenia and Pseudoleucocytosis

Hubert Schrezenmeier
1   The Department of Medicine III, University of Ulm, Ulm, Germany
,
Hilke Müller
1   The Department of Medicine III, University of Ulm, Ulm, Germany
,
Eberhard Gunsilius
1   The Department of Medicine III, University of Ulm, Ulm, Germany
,
Hermann Heimpel
1   The Department of Medicine III, University of Ulm, Ulm, Germany
,
Erhard Seifried
2   The Institute of Transfusion Medicine, Blood Donor Service Hessen, Frankfurt, Germany
› Author Affiliations
Further Information

Publication History

Received 21 January 1994

Accepted after resubmission 30 November 1994

Publication Date:
09 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Pseudothrombocytopenia (PTP) is the phenomenon of falsely low platelet counts due to in vitro platelet clumping in the presence of platelet autoantibodies and anticoagulants. We assessed anticoagulant- dependence, time course of platelet counts and impact of different counter devices on the phenomenon. Blood of 10 persons with previously recognized pronounced EDTA-dependent PTP was collected into 7 different anticoagulants and counted after different intervals in parallel in a Coulter T540 and a Coulter STKS counter and by phase contrast microscopy. With the Coulter T540 model PTP was most pronounced in blood samples anticoagulated with EDTA, Na-oxalate or Na-citrate. In the STKS counter EDTA, heparin and oxalate presented as the worst anticoagulants. The time course of platelet counts was significantly different between the two counters. Our results demonstrate that PTP is not restricted to EDTA, but is also present with other anticoagulants. In contrast, pseudoleucocytosis was observed only in EDTA-anticoagulated blood in the Coulter T540 device.

We investigated the expression of platelet integrins and activation antigens on platelets of persons with anticoagulant-dependent PTP and in healthy controls without PTP. In the presence of EDTA the expression of GpIIb/IIIa was significantly reduced in the PTP subjects compared to control. Activation antigens CD62, CD63 and thrombospondin-antigen were upregulated in the presence of EDTA. These alterations in the expression of platelet antigens could also be induced on platelets of normal donors by incubation with sera of PTP subjects and EDTA. The downregulation of GP IIb/IIIa in EDTA-blood of PTP subjects supports the hypothesis that the GPIIb/IIIa complex might be the binding site for the anti-platelet antibodies present in the sera of PTP subjects. Our results suggest that the binding of the autoantibodies causes platelet activation similar to other platelet-activating stimuli leading to subsequent platelet agglutination.

 

  • References

  • 1 Gowland E, Kay HE M, Spillman JC, Williamson JR. Agglutination of platelets by a serum factor in the presence of EDTA. J Clin Pathol 1969; 22: 460-464
    CrossrefPubMedGoogle Scholar
  • 2 Shreiner DP, Bell WR. Pseudothrombocytopenia: Manifestation of a new type of platelet agglutinin. Blood 1973; 42: 541-549
    PubMedGoogle Scholar
  • 3 Kjeldsberg CR, Swanson J. Platelet satellitism. Blood 1974; 43: 831-836
    PubMedGoogle Scholar
  • 4 Kjeldsberg CR, Hershgold EJ. Spurious thrombocytopenia. JAMA 1974; 227: 628-630
    CrossrefPubMedGoogle Scholar
  • 5 Watkins SP, Shulman NR. Platelet Cold Agglutinins. Blood 1970; 36: 153-158
    PubMedGoogle Scholar
  • 6 Lombarts AJ, de Kieviet W. Recognition and prevention of pseudothrombocytopenia and concomitant pseudoleukocytosis. Am J Clin Pathol 1988; 89: 634-639
    CrossrefPubMedGoogle Scholar
  • 7 Savage RA. Pseudoleukocytosis due to EDTA-induced platelet clumping. Am J Clin Pathol 1984; 81: 317-322
    CrossrefPubMedGoogle Scholar
  • 8 Payne BA, Pierre RV. Pseudothrombocytopenia: a laboratory artifact with potentially serious consequences. Mayo Clin Proc 1984; 59: 123-125
    CrossrefPubMedGoogle Scholar
  • 9 Vicari A, Banfi G, Bonini PA. EDTA-dependent pseudothrombocytopaenia: a 12-month epidemiological study. Scand J Clin Lab Invest 1988; 48: 537-542
    CrossrefPubMedGoogle Scholar
  • 10 Nilsson T, Norberg B. Thrombocytopenia and pseudothrombocytopenia: a clinical and laboratory problem. Scand J Haematol 1986; 37: 341-346
    PubMedGoogle Scholar
  • 11 Berkman N, Michaeli Y, Or R, Eldor A. EDTA-dependent pseudothrombocytopenia: a clinical study of 18 patients and a review of the literature. Am J Hematol 1991; 36: 195-201
    CrossrefPubMedGoogle Scholar
  • 12 Garcia SuarezJ, Calero MA, Ricard MP, Krsnik I, Rus GP, Perera F. et al EDTA-dependent pseudothrombocytopenia in ambulatory patients: clinical characteristics and role of new automated cell-counting in its detection [letter]. Am J Hematol 1992; 39: 146-148
    CrossrefPubMedGoogle Scholar
  • 13 Veenhoven WA, Van der Schans GS, Huiges W, Metting-Scherphuis HE, Halie MR, Nieweg HO. Pseudothrombocytopenia due to agglutinins. Am J Clin Pathol 1979; 72: 1005-1008
    CrossrefPubMedGoogle Scholar
  • 14 Manthorpe R, Kofod B, Wiik A, Saxtrup O, Svehag S-E. Pseudothrombocytopenia - in vitro studies on the underlying mechanism. Scand J Haematol 1981; 26: 385-392
    PubMedGoogle Scholar
  • 15 Onder O, Weinstein A, Hoyer LW. Pseudothrombocytopenia caused by platelet agglutinins that are reactive in blood anticoagulated with chelating agents. Blood 1980; 56: 177-182
    PubMedGoogle Scholar
  • 16 Mant MJ, Doery JC G, Gauldie J, Sims H. Pseudothrombocytopenia due to platelet aggregation and degranulation in blood collected in EDTA. Scand J Haematol 1975; 15: 161-170
    PubMedGoogle Scholar
  • 17 Pegels JG, Bruynes EC E, Engelfriet CP, von dem Borne AE G K. Pseudothrombocytopenia: An immunologie study on platelet antibodies dependent on ethylene diamine tetra-acetate. Blood 1982; 59: 157-161
    PubMedGoogle Scholar
  • 18 Castaman G, Ruggeri M, Rodeghiero F. EDTA-dependent pseudothrombocytopenia [letter]. Am J Hematol 1992; 41: 226
    CrossrefPubMedGoogle Scholar
  • 19 van Vliet HH, Kappers KlunneM, Abels J. Pseudothrombocytopenia: a cold autoantibody against platelet glycoprotein GP IIb. Br J Haematol 1986; 62: 501-511
    CrossrefPubMedGoogle Scholar
  • 20 Rosenkranz B, Wisser H, Bode JC. Pseudothrombocytopenia - an error in the determination of thrombocyte count. Dtsch Med Wochenschr 1985; 110: 492-495
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Forscher CA, Sussman II, Friedman EW, Solomon V, Spaet TH. Pseudothrombocytopenia masking true thrombocytopenia. Am J Hematol 1985; 18: 313-317
    CrossrefPubMedGoogle Scholar
  • 22 Solanki DL, Blackburn BC. Spurious leukocytosis and thrombocytopenia. JAMA 1983; 250: 2515-2524
    PubMedGoogle Scholar
  • 23 Berning H, Stilbo I. Pseudothrombocytopenia and the haematology laboratory. Lancet 1982; 2: 1469-1470
    PubMedGoogle Scholar
  • 24 Tschoeppe D, Schauseil S, Roesen P, Kaufmann L, Gries FA. Darstellung von Thrombozytenmembranglykoproteinen mit monoklonalen Antikörpern im durchflußzytometrischen Bioassay. Klin Wo Schr 1988; 66: 117-122
    PubMedGoogle Scholar
  • 25 Payne BA. EDTA-induced pseudothrombocytopenia. Recognizing a laboratory artifact Postgrad Med 1985; 77: 75-76
    PubMedGoogle Scholar
  • 26 Lippi U, Schinella M, Nicoli M, Modena N, Lippi G. EDTA-induced platelet aggregation can be avoided by a new anticoagulant also suitable for automated complete blood count. Haematologica 1990; 75: 38-41
    PubMedGoogle Scholar
  • 27 Boehme WM, Mahmod T, Phanuphak P. Case report. Pseudothrombocytopenia associated with vasculitis Am J Med Sci 1980; 279: 125-128
    CrossrefPubMedGoogle Scholar
  • 28 Walenga JM, Hoppensteadt D, Koza M, Wallock M, Pifarre R, Fareed J. Laboratory assays for the evaluation of recombinant hirudin. Haemostasis 1991; 21 suppl (Suppl. 01) 49-63
    PubMedGoogle Scholar
  • 29 Kettner C, Shaw E. D-Phe-Pro-ArgCH2Cl-A selective affinity label for thrombin. Thromb Res 1979; 14: 969-973
    CrossrefPubMedGoogle Scholar
  • 30 Thompson CB, Diaz DD, Quinn PG, Lapins M, Kurtz SR, Valeri CR. The role of anticoagulation in the measurement of platelet volumes. Am J Clin Path 1983; 80: 327-332
    CrossrefPubMedGoogle Scholar
  • 31 Nurden AT, Caen JP. An abnormal platelet glycoprotein pattern in three cases of GlanzmamTs thrombasthenia. Br J Haematol 1974; 28: 253-260
    CrossrefPubMedGoogle Scholar
  • 32 McGregor JL, Brochier J, Wild F, Follea G, Trzeciak M-C, James E. et al. Monoclonal antibodies against platelet membrane glycoproteins. Characterization and effect on platelet function Eur J Biochem 1983; 131: 427-436
    CrossrefPubMedGoogle Scholar
  • 33 McGregor JL, McGregor L, Bauer A-S, Catimel B, Brochier J, Dechavanne M. et al. Identification of two distinct regions within the binding sites for fibrinogen and fibronectin on the IIb-IIIa human platelet membrane glycoprotein complex by monoclonal antibodies P2 and P4. Eur J Biochem 1986; 159: 443-449
    CrossrefPubMedGoogle Scholar
  • 34 Behnke O. Surface membrane clearing of receptor-ligand complexes in human blood platelets. Journal of Cell Science 1987; 87: 465-472
    PubMedGoogle Scholar
  • 35 Wencel-Drake JD. Plasma membrane GPIIb/IIIa. Evidence for a cycling receptor pool American Journal of Pathology 1990; 136: 61-70
    PubMedGoogle Scholar
  • 36 Metzelaar MJ, Korteweg J, Sixma JJ, Nieuwenhuis HK. Comparison of platelet membrane markers for the detection of platelet activation in vitro and during platelet storage and cardiopulmonary bypass surgery. J Lab Clin Med 1993; 121: 579-587
    PubMedGoogle Scholar
  • 37 Nishibori M, Cham B, McNicol A, Shalev A, Jain N, Gerrard JM. The protein CD63 in in platelet dense granules, is deficient in a patient with Hermansky-Pudlak syndrome, and appears identical to granulophysin. Journal of Clinical Investigation 1993; 91: 1775-1782
    CrossrefPubMedGoogle Scholar
  • 38 Frazier WA. Thrombospondin: a modular adhesive glycoprotein of platelets and nucleated cells. Journal of Cellular Biology 1987; 105: 625-630
    CrossrefPubMedGoogle Scholar