Leytin Valery, Shakoor Samantha, Mody Meera, Allen David, Garvey Bernadette, Freedman John
Department of Transfusion Medicine, Shuter Wing, Room 2003, St. Michael's Hospital, 30 Bond Street, Toronto, Ontario M5B 1W8, Canada.
Crit Care Med. 2002 Dec;30(12):2771-3. doi: 10.1097/00003246-200212000-00025.
To analyze the effect of cytokines generated in sepsis and endotoxemia (tumor necrosis factor [TNF]-alpha and interleukins [IL]-1beta, -6, and -8) on activation of human platelets and to study the effect of cytokines on platelet activation in the presence of alpha-thrombin, a potent inducer of coagulation and platelet activation generated in sepsis and endotoxemia.
flow cytometric study of platelet activation induced by cytokines and/or thrombin in the whole blood and platelet-rich plasma (PRP) of healthy volunteers.
Research laboratory in a Canadian hospital.
Nine healthy volunteers recruited from laboratory staff.
Venous blood samples were obtained into acid-citrate-dextrose anticoagulant. Whole blood and PRP were diluted with appropriate buffer optimized for analyzing platelet activation by flow cytometry. TNF-alpha, IL-1beta, IL-6, and IL-8 were added to blood or PRP in concentrations ranging from 1 to 100 ng/mL and incubated for 15 mins at 37 degrees C in the presence or absence of a submaximal concentration of human alpha-thrombin (0.025 units/mL). Samples were stained with fluorescent antibodies against markers of platelet activation (P-selectin [CD62], lysosomal protein [CD63], and fibrinogen and von Willebrand factor receptors [CD41 and CD42b, respectively]) and analyzed by flow cytometry. The data obtained show that none of these cytokines trigger activation of resting platelets in whole blood or PRP and do not modulate the effect of thrombin on platelet activation as measured by quantitation of CD62, CD63, and CD42b markers on the platelet surface.
Cytokines TNF-alpha, IL-1beta, IL-6, and IL-8, which are extensively produced in sepsis and endotoxemia, do not trigger activation of resting human platelets directly or indirectly by mediating processes in white or red blood cells. The cytokines did not affect thrombin-induced platelet activation.
分析脓毒症和内毒素血症中产生的细胞因子（肿瘤坏死因子 [TNF]-α 和白细胞介素 [IL]-1β、-6 及 -8）对人血小板活化的影响，并研究在脓毒症和内毒素血症中产生的强效凝血及血小板活化诱导剂α-凝血酶存在的情况下，细胞因子对血小板活化的影响。
对健康志愿者全血和富血小板血浆（PRP）中细胞因子和/或凝血酶诱导的血小板活化进行流式细胞术研究。
加拿大一家医院的研究实验室。
从实验室工作人员中招募的9名健康志愿者。
采集静脉血样本至含酸-柠檬酸-葡萄糖抗凝剂中。全血和PRP用经优化的合适缓冲液稀释，以通过流式细胞术分析血小板活化。将TNF-α、IL-1β、IL-6和IL-8以1至100 ng/mL的浓度添加至血液或PRP中，在存在或不存在亚最大浓度人α-凝血酶（0.025单位/mL）的情况下于37℃孵育15分钟。样本用针对血小板活化标志物（P-选择素 [CD62]、溶酶体蛋白 [CD63] 以及纤维蛋白原和血管性血友病因子受体 [分别为CD41和CD42b]）的荧光抗体染色，并通过流式细胞术进行分析。所获数据显示，这些细胞因子均未触发全血或PRP中静息血小板的活化，并且如通过定量血小板表面的CD62、CD63和CD42b标志物所测，它们不会调节凝血酶对血小板活化的作用。
在脓毒症和内毒素血症中大量产生的细胞因子TNF-α、IL-1β、IL-6和IL-8，不会直接或通过介导白细胞或红细胞中的过程间接触发静息人血小板的活化。这些细胞因子不影响凝血酶诱导的血小板活化。
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