Glenister Kristen M, Payne Katherine A, Sparrow Rosemary L
Research Unit, Australian Red Cross Blood Service, Melbourne, Victoria, Australia.
Transfusion. 2008 Jan;48(1):99-107. doi: 10.1111/j.1537-2995.2007.01487.x. Epub 2007 Sep 24.
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.
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.
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.
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.
血小板（PLT）储存损伤仍未完全明确。为了更深入了解PLT储存损伤，我们对白细胞滤除的混合富血小板血浆（PC）浓缩物上清液进行了蛋白质组学分析。
PC在PLT添加剂溶液中制备，并按照标准血库程序储存。在储存的7天内收集上清液样本。通过二维（2D）凝胶电泳和质谱生成上清液蛋白质图谱。使用细胞因子抗体微阵列和酶联免疫吸附测定来研究生物活性分子。
PC上清液蛋白质的2D凝胶图谱显示出许多血浆蛋白质图谱的特征。鉴定出了几种储存诱导的蛋白质变化，包括主要血浆蛋白质的修饰。还鉴定出了PLT衍生的蛋白质，包括类震颤转录本1和整合素连接激酶，它们可能影响PLT与内皮的相互作用。细胞因子抗体微阵列揭示了许多以前与输血用PC无关的生物活性蛋白质，如脑源性神经营养因子（BDNF）。在PC储存期间，包括BDNF、CXCL7、表皮生长因子、PLT衍生生长因子（PDGF）和CCL5在内的PLT衍生细胞因子浓度显著增加。从第5天到第7天延长储存导致PC上清液中BDNF、PDGF和CCL5水平显著升高。
蛋白质组学技术为储存对PC的影响以及可溶性蛋白质对PLT储存损伤发展和受者对PLT输血反应的贡献提供了有价值的新见解。
