Thrombin generation and bleeding in haemophilia A


  • Parts of this work have been presented at the 2007 Annual National Hemophilia Foundation Meeting [abstract: Haemophilia 2008:14(4)] and the 2008 World Federation Hemophilia Meeting.

Dr Kathleen E. Brummel-Ziedins, Department of Biochemistry, University of Vermont, 208 South Park Drive, Suite 2, Colchester, VT 05446, USA.
Tel.: +1 802 656 9599; fax: +1 802 656 2256;


Summary.  Haemophilia A displays phenotypic heterogeneity with respect to clinical severity. The aim of this study was to determine if tissue factor (TF)-initiated thrombin generation profiles in whole blood in the presence of corn trypsin inhibitor (CTI) are predictive of bleeding risk in haemophilia A. We studied factor(F) VIII deficient individuals (11 mild, 4 moderate and 12 severe) with a well-characterized 5-year bleeding history that included haemarthrosis, soft tissue haematoma and annual FVIII concentrate usage. This clinical information was used to generate a bleeding score. The bleeding scores (range 0–32) were separated into three groups (bleeding score groupings: 0, 0 and ≤9.6, >9.6), with the higher bleeding tendency having a higher score. Whole blood collected by phlebotomy and contact pathway suppressed by 100 μg mL−1 CTI was stimulated to react by the addition of 5 pm TF. Reactions were quenched at 20 min by inhibitors. Thrombin generation, determined by enzyme-linked immunosorbent assay for thrombin–antithrombin was evaluated in terms of clot time (CT), maximum level (MaxL) and maximum rate (MaxR) and compared to the bleeding score. Data are shown as the mean±SD. MaxL was significantly different (< 0.001) between the groups: 504 ± 114, 315 ± 117 and 194 ± 91 nm; with higher thrombin concentrations in the groups with lower bleeding scores. MaxR was higher in the groups with a lower bleeding score; 97 ± 51, 86 ± 60 and 39 ± 16 nm min−1 (= 0.09). No significant difference was detected in CT among the groups, 5.6 ± 1.3, 4.7 ± 0.7 and 5.6 ± 1.3 min. Our empirical study in CTI-inhibited whole blood shows that the MaxL of thrombin generation appears to correlate with the bleeding phenotype of haemophilia A.