In vitro function and phagocytosis of galactosylated platelet concentrates after long-term refrigeration
In vitro function and phagocytosis of galactosylated platelet concentrates after long-term refrigeration
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DOI：
10.1111/j.1537-2995.2007.01134.x
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发表时间：
2007-03-01
期刊：
影响因子：
2.9
通讯作者：
Hoffmeister, Karin M.
中科院分区：
文献类型：
--
作者：
Babic, Aleksandar M.;Josefsson, Emma C.;Hoffmeister, Karin M.
背景：在没有血浆的情况下短期冷冻血小板（PLT）可导致其在输血后快速清除。用半乳糖阻断糖蛋白Ib α（GPIb α）的β-N-乙酰葡糖胺（β-GlcNAc）残基可防止冷藏的人和小鼠PLT与巨噬细胞结合，并缩短冷藏小鼠PLT的循环时间。PLT相关半乳糖基转移酶在其底物UDP存在的情况下有效地半乳糖基化冷冻的PLT-半乳糖被添加到富含PLT的血浆中。为了表征在血库中处理的冷藏和半乳糖基化的人PLT的止血功能，在静态和流动条件下体外研究了PLT聚集以及整合素β 3（CD 61）、CD 62 P（P-选择素）、GP 1b α（CD 42 b）、膜联蛋白V结合和整联蛋白所有3种活化。巨噬细胞对半乳糖基化冷藏血小板的亲和力用识别和吞噬冷藏血小板的THP-1细胞进行评价。冷藏和半乳糖基化14天的PLT 1）当在标准聚集测定中暴露于激动剂时保持其聚集的能力，2）与室温（RT）储存的PLT相比，显示出较不显著的GPIb α表面表达变化，3）P-选择素表达增加，和4）在体外被分化的THP-1细胞吞噬不良。此外，它表明，冷藏的血小板不影响他们的粘附性能在体外flowconditions.CONCLUSION：它表明，冷藏的人血小板保留在体外功能优于RT血小板在存储过程中，并证明，半乳糖基化防止识别存储的冷藏血小板在体外巨噬细胞。
BACKGROUND: Short-term refrigeration of platelets (PLTs) in the absence of plasma results in their rapid clearance after transfusion. Blocking beta-N-acetylglucosamine (beta-GlcNAc) residues of glycoprotein lb alpha (GPlb alpha) with galactose prevents binding of refrigerated human and mouse PLTs to macrophages and prolongs the circulation times of refrigerated mouse PLTs. PLT-associated galactosyltransferase efficiently galactosylates chilled PLTs in the presence of its substrate UDP-galactose is added to PLT-rich plasma.STUDY DESIGN AND METHODS: To characterize the hemostatic function of refrigerated and galactosylated human PLTs processed in the blood bank, PLT aggregation was studied in vitro under static and flow conditions and expression of integrin beta 3 (CD61), CD62P (P-selectin), GPlb alpha (CD42b), annexin V binding, and integrin all 3 activation with flow cytometry. Affinity of macrophages for galactosylated refrigerated PLTs was evaluated with THP-1 cells, which recognize and phagocytize refrigerated PLTs.RESULTS: PLTs refrigerated and galactosylated for 14 days 1) maintained their ability to aggregate when exposed to agonists in a standard aggregometry assay, 2) showed less pronounced changes in surface expression of GPlb alpha compared with room temperature (RT)stored PLTs, 3) increased P-selectin expression, and 4) were poorly phagocytized by differentiated THP-1 cells in vitro. In addition, it is shown that refrigeration of PLTs does not affect their adhesive properties under in vitro flow conditions.CONCLUSION: It is shown that refrigerated human PLTs retain in vitro function better than RT PLTs during storage and demonstrate that galactosylation prevents recognition of stored refrigerated PLTs by macrophages in vitro.