Aznar J A, Bonanad S, Montoro J M, Hurtado C, Cid A R, Soler M A, De Miguel A
Unidad de Coagulopatias Congénitas, Hospital Universitario La Fe, Valencia, España.
Vox Sang. 2000;79(3):156-60. doi: 10.1159/000031234.
To study the influence of virus photoinactivation with methylene blue (MB) on the coagulation factors of fresh frozen plasma (FFP) and the corresponding cryoprecipitates and cryosupernatants derived from it.
The photoinactivation procedure of the German Red Cross (Springe) was applied using Biomat (Grifols, Spain). Twenty isogroup pools of three plasma units were made from 60 U of FFP. The pools were split into three bags. One of them was photoinactivated, and pre- and postinactivation samples (MB-plasma) were obtained. The second bag was treated in the same way, followed by the preparation of MB-cryoprecipitate and MB-cryosupernatant. The third bag was not photoinactivated, and was processed in the same way to obtain control cryoprecipitate and cryosupernatant. The prothrombin time and activated partial thromboplastin time were analysed, as well as fibrinogen, factors (F) II, V, VII, VIII, IX, XI and XIII, antithrombin III, von Willebrand (vW) F:RCo, vWF:Ag and the multimeric structure of vWF.
In plasma, the proteins most sensitive to photoinactivation were fibrinogen, FV, FVIII, FIX and FXI (24, 32, 28, 23 and 27% loss, respectively). In the MB-cryoprecipitate, the losses were higher for FVIII (23%), moderate for fibrinogen, FXIII and vWF:RCo (18, 14 and 13%, respectively) and minimal (only 3%) for vWF:Ag. In MB-cryosupernatants, the losses were higher for FV (26%) and moderate for fibrinogen (16%), FIX (18%) and FXI (19%), as well as for FII and FXIII (15%). The multimeric structure of vWF was not modified in MB-plasma or in MB-cryoprecipitates. The supernatants (both MB treated as well as controls) showed an absence of multimers of very high and high molecular weight.
The quantitative and qualitative conservation of coagulation factors achieved in MB-plasma-derived products suggest that they are useful for the global replacement of coagulation factors and for deficiencies in FV and FXI. In countries lacking the economic resources to obtain virally inactivated concentrates, MB-cryoprecipitates could be useful in von Willebrand's disease and fibrinogen and FXIII deficiencies. MB-cryosupernatants could be employed in thrombotic thrombocytopenic purpura, in the correction of total or partial deficiencies of prothrombin complex factors and in specific deficiencies of FV and FXI.
研究亚甲蓝（MB）光化学病毒灭活对新鲜冰冻血浆（FFP）及其相应冷沉淀和冷上清液中凝血因子的影响。
采用西班牙Grifols公司的Biomat，应用德国红十字会（施普林格）的光化学病毒灭活程序。从60单位FFP中制备20个由三个血浆单位组成的同组混合样本。将这些混合样本分装到三个袋子中。其中一袋进行光化学病毒灭活，并获取灭活前后的样本（MB-血浆）。第二袋以同样方式处理，随后制备MB-冷沉淀和MB-冷上清液。第三袋不进行光化学病毒灭活，以同样方式处理以获取对照冷沉淀和冷上清液。分析凝血酶原时间、活化部分凝血活酶时间，以及纤维蛋白原、因子（F）II、V、VII、VIII、IX、XI和XIII、抗凝血酶III、血管性血友病因子（vW）F:RCo、vWF:Ag和vWF的多聚体结构。
在血浆中，对光化学病毒灭活最敏感的蛋白质是纤维蛋白原、FV、FVIII、FIX和FXI（分别损失24%、32%、28%、23%和27%）。在MB-冷沉淀中，FVIII损失更高（23%），纤维蛋白原、FXIII和vWF:RCo损失适中（分别为18%、14%和13%），vWF:Ag损失最小（仅3%）。在MB-冷上清液中，FV损失更高（26%），纤维蛋白原（16%）、FIX（18%）和FXI（19%）以及FII和FXIII（15%）损失适中。vWF的多聚体结构在MB-血浆或MB-冷沉淀中未发生改变。上清液（MB处理组和对照组）均未显示出高分子量和极高分子量的多聚体。
MB-血浆衍生产品中凝血因子在数量和质量上的保存表明，它们可用于全面替代凝血因子以及治疗FV和FXI缺乏症。在缺乏经济资源获取病毒灭活浓缩物的国家，MB-冷沉淀可用于治疗血管性血友病、纤维蛋白原缺乏症和FXIII缺乏症。MB-冷上清液可用于血栓性血小板减少性紫癜、纠正凝血酶原复合物因子的全部或部分缺乏症以及治疗FV和FXI的特定缺乏症。
