Journal of the Chinese Medical Association
Volume 69, Issue 12 , Pages 569-574, December 2006

Screening for Platelet Antibodies in Adult Idiopathic Thrombocytopenic Purpura: A Comparative Study Using Solid Phase Red Cell Adherence Assay and Flow Cytometry

  • Jeong-Shi Lin

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • Division of Hematology and Oncology, Department of Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • National Yang-Ming University School of Medicine, Taipei, Taiwan, R.O.C.
    • Corresponding Author InformationCorrespondence to: Dr Jeong-Shi Lin, Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, 201, Section 2, Shih-Pai Road, Taipei 112, Taiwan, R.O.C.
    • ,
  • Jau-Yi Lyou

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • ,
  • Ying-Ju Chen

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • ,
  • Pei-Shan Chen

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • ,
  • Hsueng-Mei Liu

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • ,
  • Chao-Hung Ho

      Affiliations

    • Division of Hematology and Oncology, Department of Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • National Yang-Ming University School of Medicine, Taipei, Taiwan, R.O.C.
    • ,
  • Tsung-Chi Hao

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • National Yang-Ming University School of Medicine, Taipei, Taiwan, R.O.C.
    • ,
  • Cheng-Hwai Tzeng

      Affiliations

    • Division of Transfusion Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • Division of Hematology and Oncology, Department of Medicine, Taipei Veterans General Hospital, Taiwan, R.O.C.
    • National Yang-Ming University School of Medicine, Taipei, Taiwan, R.O.C.

Received 26 May 2006; accepted 13 November 2006.

Article Outline

Background

Idiopathic thrombocytopenic purpura (ITP) is an autoimmune disorder caused by antiplatelet autoantibodies. In this study, we compared 2 methods for screening serum platelet antibodies in patients with ITP.

Methods

A total of 44 adult patients were clinically classified with ITP. We used 2 indirect tests to detect human leukocyte antigen antibodies and/or platelet-specific antibodies in their sera. In method I, we used solid phase red cell adherence (SPRCA) assay. In method II, by flow cytometry, platelets from plateletpheresis components were used as target cells, and fluorescein isothiocyanate-conjugated sheep anti-human IgG Fc was used as the staining reagent. Positive results were defined as any test with the percentage of fluorescence exceeding the reference range by 3% or more in method II. Direct tests detecting platelet-associated IgG on platelets of patients with ITP were done by flow cytometry.

Results

Serum specimens from 44 adult patients with ITP (28 female, 16 male) were tested. SPRCA assay could only detect platelet antibodies in 22 patients (50%). By method II, 31 serum specimens (70.5%) yielded positive results. There was a difference between the results of the SPRCA test and method II, with a high degree of significance (p < 0.001) by the McNemar test. No significant difference in platelet counts was observed for patients with and without discernible platelet antibodies by SPRCA assay (p = 0.90). The direct test was positive in 12 patients (66.7%) out of 18 ITP patients tested.

Conclusion

Flow cytometry is more sensitive than SPRCA assay for detecting platelet antibodies. Detection of platelet antibodies is useful in explaining the immune mechanism and platelet transfusion refractoriness in ITP.

Key Words:  flow cytometry , human leukocyte antigen , idiopathic thrombocytopenic purpura , platelet antibody , solid phase red cell adherence assay

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References 

  1. Yildirmak Y , Yanikkaya-Demirel G , Palanduz A , Kayaalp N . Antiplatelet antibodies and their correlation with clinical findings in childhood immune thrombocytopenic purpura . Acta Haematologica . 2005;113:109–112
  2. Chow MP, Yung CH, Hu HY, Tzeng JL, Lin JS, Lin WM, et al. HLA antibodies: the cause of platelet alloimmunization in Chinese . Am J Hematol . 1992;39:15–19
  3. Lo SC , Lin DT , Lin SWS , Chang JS . Frequency and characterization of platelet-specific antibodies in patients who received multiple platelet transfusions . J Formos Med Assoc . 2000;99:902–905
  4. Novotny VM , van Doorn R , Witvliet MD , Claas FH , Brand A . Occurrence of allogeneic HLA and non-HLA antibodies after transfusion of prestorage filtered platelets and red blood cells: a prospective study . Blood . 1995;85:1736–1741
  5. Kurz M, Greinix H, Hocker P, Kalhs P, Knobl P, Mayr WR, et al  Specificities of anti-platelet antibodies in multi-transfused patients with haemato-oncological disorders . Br J Haematol . 1996;95:564–569
  6. Godeau B , Fromont P , Seror T , Duedari N , Bierling P . Platelet alloimmunization after multiple transfusions: a prospective study of 50 patients . Br J Haematol . 1992;81:395–400
  7. McMillan R , Tani P , Millard F , Berchtold P , Renshaw L , Woods VL . Platelet-associated and plasma anti-glycoprotein autoantibodies in chronic ITP . Blood . 1987;70:1040–1045
  8. Tani P , Berchtold P , McMillan R . Autoantibodies in chronic ITP . Blut . 1989;59:59; 44-6.
  9. Davoren A , Bussel J , Curtis BR , Moghaddam M , Aster RH , McFarland JG . Prospective evaluation of a new platelet glycopro-tein (GP)-specific assay (PakAuto) in the diagnosis of autoimmune thrombocytopenia (AITP) . Am J Hematol . 2005;78:193–197
  10. Berchtold P , McMillan R , Tani P , Sommerville-Nielsen S , Blanchette VS . Autoantibodies against platelet membrane glyco-proteins in children with acute and chronic immune thrombo-cytopenic purpura . Blood . 1989;74:1600–1602
  11. Rachel JM , Summers TC , Sinor LT , Plapp FV . Use of a solid phase red blood cell adherence method for pretransfusion platelet compatibility testing . Am J Clin Pathol . 1988;90:63–68
  12. Jones CD , Gould LM , Lee S . An evaluation of a solid phase red cell adherence test for detecting platelet-associated IgG in immune thrombocytopenia . Am J Clin Pathol . 1990;93:552–554
  13. von dem Borne AE , Verheugt FW , Oosterhof F , von Riesz E , de la Riviere AB , Engelfriet CP . A simple immunofluorescence test for the detection of platelet antibodies . Br J Haematol . 1978;39:195–207
  14. Kiefel V , Santoso S , Weisheit M , Mueller-Eckhardt C . Monoclonal antibody-specific immobilization of platelet antigens (MAIPA): a new tool for the identification of platelet-reactive antibodies . Blood . 1987;70:1722–1726
  15. Worfold LA , MacPherson BR . The detection of platelet allo-antibodies by flow cytometry . Transfusion . 1991;31:340–344
  16. Marshall LR , Brogden FE , Roper TS , Barr AL . Antenatal platelet antibody testing by flow cytometry: results of a pilot study . Transfusion . 1994;34:961–965
  17. Helmberg W , Folsch B , Wagner T , Lanzer G . Detection and differentiation of platelet-specific antibodies by flow cytometry: the bead-mediated platelet assay . Transfusion . 1997;37:502–506
  18. Stockelberg D , Hou M , Jacobsson S , Kutti J , Wadenvik H . Detection of platelet antibodies in chronic idiopathic throm-bocytopenic purpura (ITP): a comparative study using flow cytometry, a whole platelet ELISA, and an antigen capture ELISA . Eur J Haematol . 1996;56:72–77
  19. Latorraca A, Lanza F, Moretti S, Ferrari L, Reverberi R, Galluccio L, et al. Flow cytometric analysis of anti-platelet antibodies in idiopathic thrombocytopenic purpura . Haematologica . 1994;79:269–272
  20. Romero-Guzmán LT , López-Karpovitch X , Paredes R , Barrales-Benitez O , Piedras J . Detection of platelet-associated immuno-globulins by flow cytometry for the diagnosis of immune thrombocytopenia: a prospective study and critical review . Haematologica . 2000;85:627–631
  21. George JN, Woolf SH, Raskob GE, Wasser JS, Aledort LM, Ballem PJ, et al  Idiopathic thrombocytopenic purpura: a practice guideline developed by explicit methods for the American Society of Hematology . Blood . 1996;88:3–40
  22. Bray RA , Harris SB , Josephson CD , Hillyer CD , Gebel HM . Unappreciated risk factors for transplant patients: HLA antibodies in blood components . Hum Immunol . 2004;65:240–244
  23. Tsubakio T , Tani P , Curd JG , McMillan R . Complement activation in vitro by antiplatelet antibodies in chronic immune thrombocytopenic purpura . Br J Haematol . 1986;63:293–300
  24. McMillan R , Wang L , Tomer A , Nichol J , Pistillo J . Suppression of in vitro megakaryocyte production by antiplatelet autoantibodies from adult patients with chronic ITP . Blood . 2004;103:1364–1369
  25. Pegels JG , Hermerhorst FM , van Leeuwen EF , van de Plas-van Dalen C , Engelfriet CP , von dem Borne AE . The Evans syndrome: characterization of the responsible antibodies . Br J Haematol . 1982;51:445–450
  26. Patel AP . Idiopathic autoimmune thrombocytopenia and neu-tropenia in siblings . Eur J Haematol . 2002;69:120–121
  27. Miller BA , Schultz BD . Autoimmune pancytopenia of childhood associated with multisystem disease manifestations . J Pediatr . 1983;103:877–883
  28. Jones CD , Gould LM , Lee S . An evaluation of a solid phase red cell adherence test for detecting platelet-associated IgG in immune thrombocytopenia . Am J Clin Pathol . 1990;93:552–554
  29. Raife TJ , Olson JD , Lentz SR . Platelet antibody testing in idio-pathic thrombocytopenic purpura . Blood . 1997;89:1112–1114
  30. Kelton JG , Powers P , Carter C . A prospective study of the usefulness of the measurement of platelet-associated IgG for the diagnosis of idiopathic thrombocytopenic purpura . Blood . 1982;60:1050–1053
  31. Kelton JG , Neame PB , Gauldie J , Hirsh J . Elevated platelet-associated IgG in the thrombocytopenia of septicemia . New Engl J Med . 1979;300:760–764
  32. Tavassoli M . Stress platelet: a platelet equivalent of stress retic-ulocyte . Blood Cells . 1992;18:295–300
  33. George JN , Saucerman S . Platelet IgG, IgA, IgM, and albumin: correlation of platelet and plasma concentrations in normal subjects and in patients with ITP or dysproteinemia . Blood . 1988;72:362–365

PII: S1726-4901(09)70331-4

doi:10.1016/S1726-4901(09)70331-4

Journal of the Chinese Medical Association
Volume 69, Issue 12 , Pages 569-574, December 2006

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