An in vitro analysis of the combination of hemophilia a and factor V-LEIDEN

被引:73
作者
vantVeer, C
Golden, NJ
Kalafatis, M
Simioni, P
Bertina, RM
Mann, KG
机构
[1] UNIV VERMONT,COLL MED,DEPT BIOCHEM,BURLINGTON,VT 05405
[2] UNIV PADUA,SCH MED,INST MED SEMEIOT,I-35100 PADUA,ITALY
[3] UNIV LEIDEN HOSP,CTR HEMOSTASIS & THROMBOSIS RES,NL-2300 RC LEIDEN,NETHERLANDS
关键词
D O I
10.1182/blood.V90.8.3067
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The classification of factor VIII deficiency, generally used based on plasma levels of factor VIII, consists of severe (<1% normal factor VIII activity), moderate (1% to 4% factor VIII activity), or mild (5% to 25% factor VIII activity). A recent communication described four individuals bearing identical factor VIII mutations. This resulted in a severe bleeding disorder in two patients who carried a normal factor V gene, whereas the two patients who did not display severe hemophilia were heterozygous for the factor V-LEIDEN mutation, which leads to the substitution of Arg506 --> Gln mutation in the factor V molecule. Based on the factor VIII level measured using factor VIII-deficient plasma, these two patients were classified as mild/moderate hemophiliacs. We studied the condition of moderate to severe hemophilia A combined with the factor V-LEIDEN mutation in vitro in a reconstituted model of the tissue factor pathway to thrombin. In the model, thrombin generation was initiated by relipidated tissue factor and factor VIIa in the presence of the coagulation factors X, IX, II, V, and VIII and the inhibitors tissue factor pathway inhibitor, antithrombin-III, and protein C. At 5 pmol/L initiating factor VIIa.tissue factor, a 10-fold higher peak level of thrombin formation (350 nmol/L), was observed in the system in the presence of plasma levels of factor VIII compared with reactions without factor VIII. Significant increase in thrombin formation was observed at factor VIII concentrations less than 42 pmol/L (similar to 6% of the normal factor VIII plasma concentration). In reactions without factor VIII, in which thrombin generation was downregulated by the addition of protein C and thrombomodulin, an increase of thrombin formation was observed with the factor V-LEIDEN mutation. The level of increase in thrombin generation in the hemophilia A situation was found to be dependent on the factor V-LEIDEN concentration. When the factor V-LEIDEN concentration was varied from 50% to 150% of the normal plasma concentration, the increase in thrombin generation ranged from threefold to sevenfold. The data suggested that the analysis of the factor V genotype should be accompanied by a quantitative analysis of the plasma factor V-LEIDEN level to understand the effect of factor V-LEIDEN in hemophilia A patients. The presented data support the hypothesis that the factor V-LEIDEN mutation can increase thrombin formation in severe hemophilia A. (C) 1997 by The American Society of Hematology.
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页码:3067 / 3072
页数:6
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