Proteomic analysis of supernatant from pooled buffy-coat platelet concentrates throughout 7-day storage
Proteomic analysis of supernatant from pooled buffy-coat platelet concentrates throughout 7-day storage
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DOI：
10.1111/j.1537-2995.2007.01487.x
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发表时间：
2008-01-01
期刊：
影响因子：
2.9
通讯作者：
Sparrow, Rosemary L.
中科院分区：
文献类型：
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作者：
Glenister, Kristen M.;Payne, Katherine A.;Sparrow, Rosemary L.
背景：血小板（PLT）储存损伤仍不完全了解。为了更深入地了解PLT储存损伤，从leukofiltered汇集的血沉棕黄层PLT concentrates（PC）的上清液的蛋白质组学分析进行了研究设计和方法：PC中制备的PLT添加剂溶液和储存根据标准血库程序。在整个7天储存期间收集上清液样品。通过二维（2D）凝胶电泳和质谱法生成上清液蛋白质图谱。结果：PC上清蛋白质的二维凝胶图谱显示了血浆蛋白质图谱的许多特征。确定了几种储存诱导的蛋白质变化，包括对主要血浆蛋白的修饰。还鉴定了血小板衍生蛋白，包括tremlike转录物1和整合素连接激酶，其可能影响血小板-内皮相互作用。细胞因子抗体微阵列揭示了一些生物活性蛋白，这些蛋白以前没有与用于输血的PC相关，如脑源性神经营养因子（BDNF）。血小板衍生的细胞因子，包括BDNF，CXCL 7，表皮生长因子，血小板衍生的生长因子（PDGF），和CCL 5的浓度显着增加PC的存储过程中。延长存储从第5天到第7天造成的BDNF，PDGF，和CCL 5在PC supernatural.CONCLUSION：蛋白质组学技术提供了宝贵的新的洞察存储对PC的影响和可溶性蛋白质的发展的PLT存储病变和受体对PLT输血的反应。
BACKGROUND: The platelet (PLT) storage lesion remains incompletely understood. To gain a greater insight into the PLT storage lesion, a proteomic analysis of supernatant from leukofiltered pooled buffy-coat PLT concentrates (PCs) was undertaken.STUDY DESIGN AND METHODS: PCs were prepared in PLT additive solution and stored according to standard blood bank procedures. Supernatant samples were collected throughout 7 days of storage. Maps of supernatant proteins were generated by two-dimensional (2D) gel electrophoresis and mass spectrometry. Cytokine antibody microarrays and enzyme-linked immunosorbent assay were used to investigate bioactive molecules.RESULTS: The 2D gel maps of PC supernatant proteins displayed many features of plasma protein maps. Several storage-induced protein changes were identified including modifications to major plasma proteins. PLT-derived proteins were also identified, including tremlike transcript 1 and integrin-linked kinase, which may influence PLT-endothelium interactions. Cytokine antibody microarrays revealed a number of bioactive proteins that have not been previously associated with PCs produced for transfusion, such as brain-derived neurotrophic factor (BDNF). The concentration of PLT-derived cytokines including BDNF, CXCL7, epidermal growth factor, PLT-derived growth factor (PDGF), and CCL5 significantly increased during storage of PCs. Extended storage from Day 5 to Day 7 caused significantly increased levels of BDNF, PDGF, and CCL5 in PC supernatant.CONCLUSION: Proteomic techniques provide valuable new insight into the effects of storage on PCs and the contribution of soluble proteins to the development of the PLT storage lesion and recipient responses to PLT transfusion.