ADAMTS-13 regulates platelet adhesion under flow

Boris Shenkman; Ulrich Budde; Dorothea Angerhaus; Aharon Lubetsky; Naphtali Savion; Uri Seligsohn; David Varon

doi:10.1160/TH06-01-0019

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2006; 96(02): 160-166
DOI: 10.1160/TH06-01-0019

Platelets and Blood Cells

Schattauer GmbH

ADAMTS-13 regulates platelet adhesion under flow

A new method for differentiation between inherited and acquired thrombotic thrombocytopenic purpura

Boris Shenkman

¹Institute of Thrombosis and Hemostasis, Sheba Medical Center, Tel-Hashomer, Israel

,

Ulrich Budde

²Laboratory Prof. Dr. Arndt & Partner, Hamburg, Germany

,

Dorothea Angerhaus

²Laboratory Prof. Dr. Arndt & Partner, Hamburg, Germany

,

Aharon Lubetsky

¹Institute of Thrombosis and Hemostasis, Sheba Medical Center, Tel-Hashomer, Israel

,

Naphtali Savion

⁴Goldschleger Eye Research Institute, Tel-Aviv University, Tel-Aviv, Israel

,

Uri Seligsohn

¹Institute of Thrombosis and Hemostasis, Sheba Medical Center, Tel-Hashomer, Israel

,

David Varon

³Coagulation Unit, Hadassah Hebrew University Medical Center, Jerusalem, Israel

› Author Affiliations

Abstract

Summary

ADAMTS-13 cleaves large and ultra-large vonWillebrand factor multimers normally secreted by endothelial cells. Severe deficiency of this enzyme leads to thrombotic thrombocytopenic purpura (TTP). We applied the Impact-R system [Cone and plate(let) Analyzer, CPA] to determine optimal conditions for ADAMTS-13 function, to assess it’s activity in TTP patients and to distinguish inheritedTTP (inTTP) from acquiredTTP (acTTP). The role of ADAMTS-13 in platelet adhesion under different conditions was investigated applying recombinant forms of VWF and ADAMTS-13. rVWF was first treated by rADAMTS-13 either in solution or when immobilized on the surface of the well, under static or flow conditions, in the presence or absence of BaCl₂. The resulting cleavedVWF fragments were then immobilized and served to assess type 3 von Willebrand disease whole blood platelet adhesion under flow. Maximal effect of the rADAMTS-13 (decrease of platelet adhesion in the absence compared to the presence of BaCl₂), was observed when the rVWF was pre-immobilized and the cleavage step took place under flow (85%). Mixing plasma ofTTP patients with normal blood (1:3) yielded a 1.6 to 2-fold increase of platelet adhesion under flow compared to mixing normal plasma with normal blood, at shear rate range of 1,800–2,500s⁻¹. Maximal increase of platelet adhesion was observed under 2,050s⁻¹. Under these conditions, the extent of adhesion was similarly higher in patients with inTTP and acTTP versus control [surface coverage (SC) 14.5 ± 2.8% and 14.6 ± 2.5% vs. 7.4 ± 1.7%, respectively]. ADAMTS-13 activity measured by collagen-binding test was similarly low (4.2 ± 3.8% and 3.5 ± 2.4% vs. 72.2 ± 8.0%, respectively). An inverse correlation between SC and ADAMTS-13 activity was observed in a patient with inTTP assayed eight times during plasma infusion treatment. Addition of BaCl₂ to the mixture of TTP plasma and normal whole blood yieldeda decrease in platelet adhesion in inTTP (by 51%) but not in acTTP. The lack of reduction of platelet adhesion in acTTP could presumably be due to the presence of ADAMTS-13 inhibitor in these patients. These results suggest that VWF immobilization and high shear flow yielded optimal conditions for ADAMTS-13 activity, and that introduction of BaCl₂ in the Impact-R (CPA) test may be useful for differentiation between inherited and acquired TTP.

Keywords

ADAMTS-13 - TTP - VWF - Impact-R

Full Text

References

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