The basis examination of leukocyte-platelet aggregates with CD45 gating as a novel platelet activation marker
The basis examination of leukocyte-platelet aggregates with CD45 gating as a novel platelet activation marker
复制标题
CD45门控作为新型血小板活化标志物白细胞-血小板聚集体的基础检查
DOI：
10.1111/ijlh.12051
复制
发表时间：
2013
影响因子：
3
通讯作者：
Takanori Moriyama
中科院分区：
文献类型：
--
作者：
Ayumi Nagasawa;Kazuhiko Matsuno;Shogo Tamura;Koji Hayasaka;Chikara Shimizu;Takanori Moriyama
血液循环中的血小板活化被认为与血栓和炎症有关；因此，敏感和易于使用的标记是必要的。在这项研究中，我们建立了一个简单而快速的方案来临床检查白细胞-血小板聚集形成与活化血小板在循环中。方法用PC5偶联抗CD45单克隆抗体和异硫氰酸酯偶联抗CD41单克隆抗体对全血进行染色，进行白细胞-血小板聚集分析。为活化血小板，加入5 μ凝血酶受体活化肽（TRAP）或2 μ /mL胶原。样品采用EPICS XL （Beckman Coulter, Miami， FL， USA）进行分析。根据CD45荧光强度和侧散射的差异对单核细胞、中性粒细胞和淋巴细胞进行门控。对于每个门，分析表达CD41的血小板百分比（%）。用抗CD62P单克隆抗体对同一幅画进行染色。然后用血小板细胞群门控法分析血小板CD62P的表达。结果我们分析了18例健康人白细胞-血小板聚集和血小板CD62P的表达。当血小板被血小板激动剂激活时，白细胞-血小板聚集物，主要是单核细胞-血小板聚集物增加。单核细胞-血小板聚集和中性粒细胞-血小板聚集也随着时间的推移而增加，伴有轻度血小板活化。结论白细胞-血小板聚集物（主要是单核细胞-血小板聚集物）是血液循环中血小板活化的敏感标志。
IntroductionPlatelet activation in circulation is considered to be associated with thrombosis and inflammation; thus, sensitive and easy‐to‐use markers are necessary. In this study, we established a simple and rapid protocol to clinically examine leukocyte–platelet aggregate formation associated with activated platelets in circulation.MethodsWhole blood was stained with PC5‐conjugated anti‐CD45 monoclonal antibody and fluorescent isothiocyanate‐conjugated anti‐CD41 monoclonal antibody for leukocyte–platelet aggregate analysis. For platelet activation, 5 μmthrombin receptor–activated peptide (TRAP) or 2 μg/mL collagen was added. Samples were analyzed by EPICS XL (Beckman Coulter, Miami, FL, USA). Monocytes, neutrophils, and lymphocytes were gated based on differences in CD45 fluorescence intensity and side scatter. For each gate, the percentage (%) of platelets expressing CD41 was analyzed. Same drawing sample was stained with anti‐CD62P monoclonal antibody. Platelet CD62P expression was then analyzed with gating for platelet cell population.ResultsWe analyzed leukocyte–platelet aggregates and platelet CD62P expression in 18 healthy individuals. Leukocyte–platelet aggregates, mainly monocyte–platelet aggregates, increased when platelets were activated by platelet agonists. Monocyte–platelet aggregates and neutrophil–platelet aggregates also increased over time with mild platelet activation.ConclusionLeukocyte–platelet aggregates, mainly monocyte–platelet aggregates, appear to be a sensitive marker of platelet activation in circulation.