Skripchenko Andrey, Kurtz James, Moroff Gary, Wagner Stephen J
The American Red Cross Biomedical Services, Holland Laboratory, Blood Components Development, Rockville, Maryland 20855, USA.
Transfusion. 2008 Jul;48(7):1469-77. doi: 10.1111/j.1537-2995.2008.01733.x. Epub 2008 May 13.
The activation marker CD40 ligand (CD40L) has recently been demonstrated to be released from the cytoplasm of platelets (PLTs) during storage. CD40L may be associated with some adverse transfusion reactions including febrile responses and transfusion-related acute lung injury. CD62P has been traditionally measured to assess PLT activation. This study compares the surface levels of CD40L and CD62P and accumulation of the soluble forms of these activation markers in the plasma of stored PLTs, prepared by PLT-rich plasma (PRP) or buffy coat (BC) methods and with two apheresis instruments.
Individual PLT concentrates (PCs) were prepared in 100 percent plasma from a pool of two ABO-identical whole-blood units. Apheresis PLTs (APs) were prepared in 100 percent plasma using one of two commercially available cell separators (Amicus, Baxter Healthcare; and Trima, Gambro BCT). Surface expression of CD40L and CD62P was measured by flow cytometry, and secretion of soluble CD40L (sCD40L) and soluble CD62 (sCD62) was measured by enzyme-linked immunosorbent assay during 7 days of PLT storage.
Secretion of sCD40L was greater in Amicus APs than in Trima APs during the first 3 days of storage. It was also greater in the PRP-PC preparations than in BC-PC preparations through the first day of storage. Surface expression of CD40L was low in all PLT preparations. Secretion of sCD62P was greater in Amicus APs than in Trima APs during the entire storage period and greater in PRP-PC than in BC during the first 5 days of storage. The percentage of CD62P-positive PLTs was greater in Amicus units than Trima units and greater in PRP-PC than BC-PC preparations during the first 5 and 3 days of storage, respectively.
The kinetics of the secretion of CD40L are influenced by the method used to prepare PLTs for storage. The patterns for CD40L membrane association and secretion are different than those observed for CD62P during storage.
活化标志物CD40配体（CD40L）最近被证明在储存期间会从血小板（PLT）的细胞质中释放出来。CD40L可能与一些不良输血反应有关，包括发热反应和输血相关的急性肺损伤。传统上通过检测CD62P来评估血小板活化。本研究比较了通过富血小板血浆（PRP）或白膜层（BC）方法以及两种血液成分分离仪制备的储存血小板血浆中CD40L和CD62P的表面水平以及这些活化标志物可溶性形式的积累情况。
从两单位ABO血型相同的全血混合液中在100%血浆中制备单个血小板浓缩物（PC）。使用两种市售细胞分离仪（Amicus，百特医疗；和Trima，金宝BCT）之一在100%血浆中制备单采血小板（AP）。在血小板储存7天期间，通过流式细胞术检测CD40L和CD62P的表面表达，并通过酶联免疫吸附测定法检测可溶性CD40L（sCD40L）和可溶性CD62（sCD62）的分泌情况。
在储存的前3天，Amicus单采血小板中sCD40L的分泌量高于Trima单采血小板。在储存的第一天，PRP-PC制剂中sCD40L的分泌量也高于BC-PC制剂。所有血小板制剂中CD40L的表面表达均较低。在整个储存期间，Amicus单采血小板中sCD62P的分泌量高于Trima单采血小板，在储存的前5天，PRP-PC中sCD62P的分泌量高于BC。在储存的前5天和前3天，CD62P阳性血小板的百分比在Amicus单位中高于Trima单位，在PRP-PC制剂中高于BC-PC制剂。
CD40L分泌的动力学受用于制备储存血小板的方法影响。CD40L膜结合和分泌的模式与储存期间观察到的CD62P的模式不同。
