Farrugia A, Tan Y, Romeo A, Martin L, Rolland J R, Kellner S, Piouronowski H
Red Cross Blood Transfusion Service, South Melbourne, Australia.
Vox Sang. 1994;66(3):153-60. doi: 10.1111/j.1423-0410.1994.tb00302.x.
Flow cytometry was used to: (1) determine residual leucocyte numbers in red cell suspensions following the range of leucocyte depletion procedures used in our organisation, and (2) to characterize phenotypically the leucocytes using direct immunofluorescence with monoclonal antibodies to cell surface receptors. Under the conditions used, a lower limit of detection of 2.5 leucocytes per microliter (equivalent to 3.43 log10 or 99.96% removal) could be achieved. Filtration through polyester filters was found to remove up to > 99.96% of the initial leucocytes; however, a significant differential efficacy was observed between filters from different manufacturers even when filters with similar costs were compared. The order of filter brands with respect to leucocyte removal found was Pall BPF4 = Erypur Optima G-O > Sepacell R500 > Pall RC50. Phenotyping revealed that increasing filtration efficacy was associated with a preferential removal of lymphocytes; conversely, a second filtration over one brand of filter allowed proportionately more lymphocytes to pass through compared with the first filtration. A saline wash following filtration removed a further 0.5% of the initial leucocyte content, and was associated with a preferential loss of granulocytes. Freeze-thawing the red cell suspension removed fewer leucocytes (96.3%) than did filtration (98.74% to > 99.6%) or filtration followed by washing (99.22%), and also led to preferential loss of granulocytes. Flow cytometry provides a reliable tool for the quality control of leuco-depleted red cells, and allows a qualitative assessment of the residual leucocytes. This information is of value in choosing procedures aimed at decreasing the risk of alloimmunisation and post-transfusion reactions.
(1) 测定在我们机构所采用的一系列白细胞去除程序后红细胞悬液中的残留白细胞数量，以及 (2) 使用针对细胞表面受体的单克隆抗体通过直接免疫荧光对白细胞进行表型分析。在所采用的条件下，每微升可实现 2.5 个白细胞的检测下限（相当于 3.43 log10 或 99.96% 的去除率）。发现通过聚酯滤器过滤可去除高达 > 99.96% 的初始白细胞；然而，即使比较成本相似的滤器，不同制造商的滤器之间也观察到显著的差异功效。所发现的关于白细胞去除的滤器品牌顺序为 Pall BPF4 = Erypur Optima G - O > Sepacell R500 > Pall RC50。表型分析显示，过滤功效的提高与淋巴细胞的优先去除相关；相反，在一个品牌的滤器上进行第二次过滤时，与第一次过滤相比，有比例更多的淋巴细胞能够通过。过滤后用盐水洗涤可进一步去除初始白细胞含量的 0.5%，并且与粒细胞的优先损失相关。对红细胞悬液进行冻融去除的白细胞（96.3%）比过滤（98.74% 至 > 99.6%）或过滤后洗涤（99.22%）少，并且也导致粒细胞的优先损失。流式细胞术为白细胞去除红细胞的质量控制提供了一种可靠的工具，并允许对残留白细胞进行定性评估。该信息对于选择旨在降低同种免疫和输血后反应风险的程序具有价值。
