Institut für Immunologie und Transfusionsmedizin, Universitätsmedizin Greifswald, Germany.
J Proteomics. 2012 Dec 5;76 Spec No.:297-315. doi: 10.1016/j.jprot.2012.04.057. Epub 2012 May 24.
Production and storage of platelet concentrates (PC) induce protein changes in platelets leading to impaired platelet function. This study aimed to identify signaling pathways involved in the development of early platelet storage lesions in apheresis-PCs stored in plasma or additive solution (PAS). Apheresis-PCs from four donors were stored in plasma or in PAS at 22°C (n=4 each). Platelets were analyzed at day 0 (production day) and after 1, 6 and 9 days of storage. Platelet response to agonists (TRAP, collagen, ADP) and to hypotonic shock decreased, CD62P expression increased in both storage media over time. Using DIGE 1550 protein spots were monitored and compared to baseline values at day 0. Platelets in plasma displayed changes in 352 spots (166/day 1, 263/day 6 and 201/day 9); in PAS 325 spots changed (202/day 1, 221/day 6, 200/day 9). LC-ESI-MS/MS analysis of 405 platelet proteins revealed 32 proteins changed during storage in plasma (9/day 1, 15/day 6 and 26/day 9) and 28 in PAS (5/day 1, 20/day 6, 26/day 9). Ingenuity pathway analysis found integrin-αII(b)β(3) and focal adhesion signaling pathways involved in early alterations, being confirmed by Western blotting. Corresponding mRNAs in platelets were identified by next generation sequencing for 84 changed proteins. Integrin-αII(b)β(3) and focal adhesion signaling cause irreversible early storage lesions in apheresis platelets. This article is part of a Special Issue entitled: Integrated omics.
血小板浓缩物（PC）的生产和储存会导致血小板中的蛋白质发生变化，从而使血小板功能受损。本研究旨在鉴定与在血浆或添加剂溶液（PAS）中储存的机采 PC 早期储存损伤发展相关的信号通路。从四个供体中采集的机采 PC 在血浆或 PAS 中于 22°C 下储存（各储存 4 份）。在第 0 天（生产日）以及储存 1、6 和 9 天后对血小板进行分析。血小板对激动剂（TRAP、胶原、ADP）和低渗休克的反应以及 CD62P 的表达随着时间的推移在两种储存介质中均逐渐降低。使用 DIGE 监测了 1550 个蛋白质点，并与第 0 天的基线值进行了比较。在血浆中，血小板在第 1 天有 352 个点发生变化（每天 166 个，第 6 天 263 个，第 9 天 201 个）；在 PAS 中，有 325 个点发生变化（第 1 天 202 个，第 6 天 221 个，第 9 天 200 个）。对 405 个血小板蛋白质进行 LC-ESI-MS/MS 分析显示，在血浆中储存期间有 32 个蛋白质发生变化（第 1 天 9 个，第 6 天 15 个，第 9 天 26 个），在 PAS 中储存期间有 28 个蛋白质发生变化（第 1 天 5 个，第 6 天 20 个，第 9 天 26 个）。通过 Ingenuity 通路分析发现整合素-αII(b)β(3)和粘着斑信号通路参与了早期改变，这一结果通过 Western blot 得到了证实。通过下一代测序鉴定了血小板中 84 个发生变化的蛋白质对应的 mRNA。整合素-αII(b)β(3)和粘着斑信号通路导致机采血小板中不可逆的早期储存损伤。本文是一个特刊的一部分，标题为：综合组学。
