Nishino M, Miura S, Yoshioka A, Kuwahara I, Nishimura T, Hamada K, Fukui H
Nara Medical College, Department of Pediatrics, Japan.
Int J Hematol. 1993 Apr;57(2):163-73.
This is the first case of variant von Willebrand disease (vWD) with defective binding of von Willebrand factor (vWF) to factor VIII (F.VIII) to be diagnosed in Japan. An 8-year-old Japanese girl, who had had recurrent episodes of subcutaneous hematomas, showed a prolonged A-PTT, low F.VIII (F.VIII:C 4 U/dl, FVIII:Ag 4 U/dl), and normal level of vWF (RCof 80 U/dl, vWF:Ag 60 U/dl). The patient's vWF-multimeric structure on SDS agarose gel electrophoresis was similar to that in normal subjects. A F.VIII binding assay was performed, as described by Nishino et al. (1989). F.VIII binding (y) of vWF was expressed as a function of the amount of immobilized vWF (x) on the wells of a polystyrene plate. Regression lines from normal subjects and the patient had a high correlation coefficient. F.VIII binding capacity was estimated by the slope of the regression lines. The slope for normal subjects showed y = 0.002 + 0.653x, while, in contrast, the slope for the patient showed y = 0.005 + 0.009x, indicating that the capacity of vWF from the patient to bind F.VIII was markedly decreased. Exons 18-20 of the vWF gene, covering the first 132 amino acids of mature vWF subunit from the patient, were sequenced, using the PCR amplification method. A point mutation C-->T at codon 816 in exon 19, predicting a substitution of Trp for Arg(53), was characterized; this was inherited by the patient from her mother.
这是日本诊断出的首例血管性血友病因子(vWF)与凝血因子VIII(F.VIII)结合缺陷的血管性血友病变异型(vWD)病例。一名8岁日本女孩反复出现皮下血肿,活化部分凝血活酶时间(A-PTT)延长,F.VIII水平低(F.VIII:C 4 U/dl,FVIII:Ag 4 U/dl),vWF水平正常(瑞斯托霉素辅因子活性[RCof] 80 U/dl,vWF:Ag 60 U/dl)。患者vWF在十二烷基硫酸钠琼脂糖凝胶电泳上的多聚体结构与正常受试者相似。按照西野等人(1989年)所述进行F.VIII结合试验。vWF的F.VIII结合(y)表示为聚苯乙烯板孔上固定化vWF量(x)的函数。正常受试者和患者的回归线具有较高的相关系数。通过回归线的斜率估计F.VIII结合能力。正常受试者的斜率显示为y = 0.002 + 0.653x,而相比之下,患者的斜率显示为y = 0.005 + 0.009x,表明患者的vWF结合F.VIII的能力明显降低。使用聚合酶链反应(PCR)扩增方法对患者vWF基因的第18至20外显子进行测序,该外显子涵盖成熟vWF亚基的前132个氨基酸。在外显子19的第816密码子处鉴定出一个点突变C→T,预测色氨酸取代精氨酸(53);这是患者从其母亲那里遗传而来的。