de Wildt-Eggen J, Schrijver J G, Kuiper-Kramer P A, Bins M, van Prooijen H C
Bloodbank IJssellanden Zwolle, University Hospital, Utrecht, The Netherlands.
Vox Sang. 1999;77(2):97-102. doi: 10.1159/000031083.
For preparation of buffy coat-depleted red cell concentrates (RCCs) in additive solution whole blood is currently collected in The Netherlands both in quadruple-bag and bottom and top bag systems. By using the quadruple-bag system both plasma and buffy coat cells are transferred into integrated satellite bags while the red cells remain in the collection bag. When bottom and top bags are used, the buffy coat remains in the collection bag while both red cells and plasma are transferred into satellite bags. The difference in processing prompted us to perform quantitative analysis of residual WBC subsets in buffy coat-depleted RCCs. Differences in removal of specific cells with the buffy coat could improve the outcome of additional filtration procedures aiming at complete removal of specific WBC subsets.
The buffy coat was removed in semiautomated procedures (Optipress I; Compomat G4) from units of whole blood collected in both bag systems. Paired samples were taken before and after removal of the buffy coat for counting and analyzing WBC subsets by flow cytometry using subset-specific monoclonal antibodies.
All RCCs met the criteria from the guidelines of the Council of Europe. The percentage of residual total WBCs was lower (p<0.001) in RCCs processed in bottom and top bag systems (26% Compomat and 18% Optipress) as compared to RCCs processed in quadruple-bag systems (43% Compomat). WBC subset analysis in RCCs processed in quadruple-bag systems showed approximately 25% of residual T cells, B cells and monocytes and 60% of residual granulocytes. In contrast, WBC subset analysis in RCCs processed in bottom and top bag systems showed approximately 2% residual T cells, B cells, and monocytes and 35% residual granulocytes; in about 45% of units, lymphocytes and monocytes were even below the detection limit of flow cytometry analysis.
Buffy coat-depleted RCCs are currently processed in bottom and top bag or quadruple-bag systems, the former being superior to the latter due to selective depletion of lymphocytes and monocytes by 98% (2 logs). The bottom and top procedure is an evident contribution to leukodepletion in blood transfusion, both with and without additional filtration.
为制备去除白细胞层的红细胞浓缩液（RCC），目前荷兰采用添加剂溶液采集全血，使用四联袋系统以及底袋和顶袋系统。使用四联袋系统时，血浆和白细胞层细胞被转移至整合的卫星袋中，而红细胞则留在采集袋中。使用底袋和顶袋时，白细胞层留在采集袋中，红细胞和血浆都被转移至卫星袋中。这种处理方式的差异促使我们对去除白细胞层的RCC中残留白细胞亚群进行定量分析。白细胞层对特定细胞的去除差异可能会改善旨在完全去除特定白细胞亚群的额外过滤程序的效果。
采用半自动程序（Optipress I；Compomat G4）从两种袋系统采集的全血单位中去除白细胞层。在去除白细胞层前后采集配对样本，使用亚群特异性单克隆抗体通过流式细胞术计数和分析白细胞亚群。
所有RCC均符合欧洲委员会指南的标准。与四联袋系统处理的RCC（Compomat为43%）相比，底袋和顶袋系统处理的RCC中残留总白细胞的百分比更低（p<0.001）（Compomat为26%，Optipress为18%）。四联袋系统处理的RCC中的白细胞亚群分析显示，约25%为残留T细胞、B细胞和单核细胞，60%为残留粒细胞。相比之下，底袋和顶袋系统处理的RCC中的白细胞亚群分析显示，约2%为残留T细胞、B细胞和单核细胞，35%为残留粒细胞；在约45%的单位中，淋巴细胞和单核细胞甚至低于流式细胞术分析的检测限。
目前去除白细胞层的RCC采用底袋和顶袋或四联袋系统进行处理，前者由于淋巴细胞和单核细胞选择性减少98%（2个对数）而优于后者。无论有无额外过滤，底袋和顶袋程序对输血中的白细胞去除都有显著贡献。
