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单采血浆中血管性血友病因子的含量及功能活性

Content and functional activity of von Willebrand factor in apheresis plasma.

作者信息

Burnouf T, Caron C, Burkhardt T, Goudemand J

机构信息

Human Plasma Product Services, Lille, France.

出版信息

Vox Sang. 2004 Jul;87(1):27-33. doi: 10.1111/j.1423-0410.2004.00535.x.

Abstract

BACKGROUND AND OBJECTIVES

Von Willebrand Factor (VWF) is a complex high-molecular-weight (HMW) plasma glycoprotein playing a critical role in primary and secondary haemostasis. Owing to its multimeric structure and sensitivity to proteolysis, VWF can be used as a marker of the impact of collection procedures on the characteristics of plasma for transfusion and for fractionation. We studied VWF content, functional activity and HMW multimers in plasmas collected by five different automated apheresis collection procedures.

MATERIALS AND METHODS

Five series of 30 plasma units were obtained from volunteer donors at two collection sites using Haemonetics PCS2 machines with Revision (Rev) F, Rev G, high-separation core (HSC), or filter core (FC) procedures, or Baxter-Fenwall Autopheresis-C (Auto-C). VWF antigen (VWF:Ag), ristocetin cofactor (VWF:RCo) activity and HMW multimers were first determined in 10 randomly selected plasma donations collected with Rev G, HSC, FC and Auto-C procedures. Then, the same analyses and the collagen binding (VWF:CB) activity were determined in the pools of 30 donations from each of the five procedures and compared with two normal plasma pools (NPP1 and NPP2). A reference plasma (RP) was used to calibrate each assay.

RESULTS

There were a greater number of group O individuals in the Rev F, Rev G and FC donors than in the HSC and Auto-C donors. The mean VWF:Ag level was > 100 IU/dl, VWF:RCo activity was > 90 U/dl, the VWF:RCo/Ag ratio was close to 1, and the percentage of 11-15 mers was above 100% of RP in the 10 individual plasma units from Rev G, HSC, FC, and Auto-C and in their respective pools. The mean percentage of multimers > 15 mers, relative to RP, was significantly less in Rev G plasmas (48 +/- 17%; range 32-91%), compared with Auto-C, HSC and FC plasmas (P = 0.0211; 0.0257; and 0.0376, respectively). The VWF:CB activity of the 30-donation pools was 61 and 60 U/dl in Auto C and HSC, 50 U/dl in Rev F and FC, and 43 U/dl in the Rev G pool. The VWF:CB/Ag ratio was 0.54 (Auto-C), 0.49 (HSC), 0.46 (Rev F), 0.45 (FC) and 0.37 (Rev G), compared with 0.81and 0.92 in NPPs. The percentage of VWF multimers of 11-15 mers in apheresis plasma and NPP was normal. VWF multimers > 15 mers ranged from 38 to 64% of that of RP plasma, and was 111 and 112% in NPPs.

CONCLUSIONS

The VWF:Ag, VWF:RCo activity and 11-15 mer VWF multimers were well preserved in all plasma units from each of the five apheresis procedures. The VWF:CB activity and the percentage of multimers > 15 mers in apheresis plasma was less than in normal plasma pools and differed slightly among procedures.

摘要

背景与目的

血管性血友病因子(VWF)是一种复杂的高分子量(HMW)血浆糖蛋白,在原发性和继发性止血中起关键作用。由于其多聚体结构和对蛋白水解的敏感性,VWF可作为采集程序对输血和分馏用血浆特性影响的标志物。我们研究了通过五种不同的自动单采血浆采集程序采集的血浆中VWF的含量、功能活性和HMW多聚体。

材料与方法

在两个采集地点,使用配备修订版(Rev)F、Rev G、高分离核心(HSC)或滤芯(FC)程序的Haemonetics PCS2机器,或百特-芬华自动单采血浆机(Auto-C),从志愿献血者中获得五组各30个血浆单位。首先在随机选择的10份采用Rev G、HSC、FC和Auto-C程序采集的血浆捐献样本中测定VWF抗原(VWF:Ag)、瑞斯托霉素辅因子(VWF:RCo)活性和HMW多聚体。然后,对这五种程序各自的30份捐献样本混合后的血浆进行相同分析,并测定胶原结合(VWF:CB)活性,同时与两个正常血浆库(NPP1和NPP2)进行比较。使用一份参考血浆(RP)对每种检测进行校准。

结果

Rev F、Rev G和FC程序的献血者中O型个体数量多于HSC和Auto-C程序的献血者。在采用Rev G、HSC、FC和Auto-C程序采集的10份个体血浆样本及其各自的混合样本中,VWF:Ag平均水平>100 IU/dl,VWF:RCo活性>90 U/dl,VWF:RCo/Ag比值接近1,11 - 15聚体的百分比高于RP的100%。与Auto-C、HSC和FC血浆相比,Rev G血浆中多聚体>15聚体的平均百分比相对于RP显著更低(48±17%;范围32 - 91%)(分别为P = 0.0211;0.0257;和0.0376)。30份捐献样本混合后的血浆中,Auto C和HSC的VWF:CB活性为61和60 U/dl,Rev F和FC为50 U/dl,Rev G混合样本为43 U/dl。VWF:CB/Ag比值在Auto-C中为0.54,HSC中为0.49,Rev F中为0.46,FC中为0.45,Rev G中为0.37,而NPPs中为0.81和0.92。单采血浆和NPP中11 - 15聚体的VWF多聚体百分比正常。多聚体>15聚体的VWF占RP血浆的比例在38%至64%之间,而NPPs中为111%和112%。

结论

在五种单采程序各自采集的所有血浆样本中,VWF:Ag、VWF:RCo活性和11 - 15聚体的VWF多聚体均保存良好。单采血浆中VWF:CB活性和多聚体>15聚体的百分比低于正常血浆库,且不同程序之间略有差异。

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