Evaluation of von Willebrand factor-cleaving protease activity in patients with thrombotic thrombocytopenic purpura

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Journal Title:
Volume 117, Issue 06, 2004
Key Word:
von Willebrand factor;cleaving protease;collagen binding assay;thrombotic thrombocytopenic purpura;tumor

Abstract: Background Thrombotic thrombocytopenic purpura (TTP) is a rare thrombotic microangiopathy. In this study we investigated the von Willebrand factor-cleaving protease (vWF-cp) activity deficiency in patients with TTP.Methods The plasma or serum vWF-cp activity was measured using a sensitive enzyme-linked immunosorbent assay (ELISA) by detecting the residual collagen binding activity (R-CBA) of von Willebrand factor (vWF) before and after digestion by vWF-cp. Multimers of vWF in plasma of patients with TTP were also analyzed by SDS-agarose electrophoresis. Moreover, the serum vWF-cp activities were compared between the patients with TTP and those with tumors.Results The coefficient of variation for intra-batch and inter-batch of the assay were 3.60% and 8.35%. The plasma and serum vWF-cp activity in healthy individuals were (78.79±9.17)% (n=30) and (79.47±10.78)% (n=53), respectively, while the plasma vWF-cp activity in 5 patients with TTP was markedly decreased [(21.83±19.98)%, P<0.001]. The unusually large vWF multimers were observed in two plasma samples of the patients with TTP. Although the vWF-cp activities in patients with benign and malignant tumors were also decreased (P<0.03 and P<0.001, respectively), they were relatively high in comparison with that of TTP patients (P<0.001).Conclusion Measurement of the vWF-cp activity using R-CBA is a simple and rapid method for diagnosing TTP. The vWF-cp activity in patients with TTP was markedly lower than those of patients with tumors.

  • [1]Furlan M, Robles R, Galbusera M, et al. von Willebrand factor-cleaving protease in thrombotic thrombocytopenic purpura and the hemolytic-uremic syndrome. N Engl J Med 1998;339:1578-1584.
  • [2]Tsai HM, Lian EC. Antibodies to von Willebrand factor-cleaving protease in acute thrombotic thrombocytopenic purpura. N Engl J Med 1998;339:1585-1594.
  • [3]Zheng X, Chung D, Takayama TK, et al. Structure of von Willebrand factor cleaving protease (ADAMTS13), a metalloprotease involved in thrombotic thrombocytopenic purpura. J Biol Chem 2001;276:41059-41063.
  • [4]Oleksowicz L, Bhagwati N, DeLeon-Fernandez, et al. Dificient activity of von Willebrand's factor-cleaving protease in patients with disseminated malignancies. Cancer Res 1999;59:2244-2250.
  • [5]Mannucci PM, Karimi M, Mosalaei A, et al. Patients with localized and disseminated tumors have reduced but measurable levels of ADAMTS13 (von Willebrand factor cleaving protease). Haematologica 2003;88:454-458.
  • [6]Xi X, Ruan C, Li P, et al. Use of monoclonal antibodies in enzyme-linked immunoassay for von Willebrand factor. Chin J Med Lab Sci 1988;11:272-275.
  • [7]Rick M, Moll S, Taylor M, et al. Clinical use of a rapid collagen binding assay for von Willebrand factor cleaving protease in patients with thrombotic thrombocytopenic purpura. Thromb Haemost 2002;88:598-604.
  • [8]Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor Ⅶ/von Willebrand factor. Blood 1981;57:1140-1143.
  • [9]Levy GG, Nichols WC, Lian EC, et al. Mutations in a member of the ADAMTS gene family cause thrombotic thrombocytopenic purpura. Nature 2001;413:488-494.
  • [10]Moake JL. Thrombotic microangiopathies. N Engl J Med 2002;347:589-600.
  • [11]Ruggeri ZM. Old concepts and new developments in the study of platelet aggregation. J Clin Invest 2000;105:699-701.
  • [12]Studt JD, Bhm M, Budde U, et al. Measurement of von Willebrand factor-cleaving protease (ADAMTS13) activity in plasma: a multicenter comparison of different assay methods. J Thromb Haemost 2003;1:1882-1887.
  • [13]Furlan M, Lammle B. Assays of von Willebrand factor-cleaving protease: a test for diagnosis of familial and acquired thrombotic thrombocytopenic purpura. Semin Thromb Hemost 2002;28:167-172.
  • [14]Barbot J, Costa E, Guerra M, et al. Ten years of prophylactic treatment with fresh-frozen plasma in a child with chronic relapsing thrombotic thrombocytopenic purpura as a result of a congenital deficiency of von Willebrand factor-cleaving protease. Br J Haematol 2001;113:649-651.
  • [15]Tsai HM. High titers of inhibitors of von Willebrand factor-cleaving metalloproteinase in a fatal case of acute thrombotic thrombocytopenic purpura. Am J Hematol 2000;65:251-255.
  • [16]Rock G, Porta C, Bobbio-Pallavicini E, et al. Thrombotic thrombocytopenic purpura treatment in year 2000. Haematologica 2000;85:410-419.
  • [17]Blot E, Decaudin D, Veyradier A, et al. Cancer-related thrombotic microangiopathy secondary to Von Willebrand factor-cleaving protease deficiency. Thromb Res 2002;106:127-130.
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