Regulation of neutrophil functions by platelets

（1）我们已经开发了一种新的设备，它可以敏感地检测到整个血液中的平台聚合物和平台-中性聚合物的存在。脉冲激光器和CCD摄像机附着在流动细胞仪上，它是自动视网膜分析仪，R-3000（Sysmex， Japan）。前向散射的浓度和荧光强度，用于维持RNA和DNA的染料，用于选择性的门平台聚合物和平台-中性质聚合物。进入门区域的细胞被检查以正确地分配这些细胞组件的图像;通过ADP或血清素刺激，用这个新设备清楚地检测到了平台-神经质聚合物的形成。现在，我们正在努力开发一个使之能够量化测量的系统。（2）我们研究了周边血细胞的鞘脂代谢，i.e.，平台、神经细胞、神经细胞、单核细胞和单核细胞。这些高度分化的细胞中的鞘磷脂代谢的分化特征是它们的高鞘蛋白（Sph）激酶活性。大约40%的platelet Sph 1-磷酸盐（Sph-1-P）可以通过12-O-四十二anoylphorbol 13-乙酸盐释放提取物，可能通过蛋白激酶C介导。在另一只手上，在细胞、中性植物和单核细胞中，Sph-1-P形成了细胞内部的显著百分比，而在没有刺激的情况下，刺激依赖释放是边际的。Sphh及其甲基化衍生物， N，N-二甲基蛋白酶，诱导的凋亡不只是在中性植物中，但也在单核细胞中，而Sph-l-P eliced CaïD 22 + YïD 2在平台中的移动。我们的结果建议所有的血液细胞都可以去除等离子体Sphh，哪些是有害的或抑制的，并将其转换为Sph-1-P，作为等离子体Sph-1-P的来源，哪些可能在血液容器中扮演各种重要角色？

(1) We have developed a new device which can sensitively detect the presence of platelet aggregates and platelet-neutrophil aggregates in whole blood. A pulse laser and a CCD camera are attached to a flow cytometer, which is an automated reticulocyte analyzer, R-3000 (Sysmex, Japan). Forward scatter intensity and fluorescence intensity of auramine O, a dye for staining RNA and DNA, are used to selectively gate platelet aggregates and platelet-neutrophil aggregates. Cells that fall in the gated area are checked for their images to correctly distinguish these aggregates from other cell components.Upon stimulation with ADP or serotonin, formation of platelet-neutrophil aggregates was clearly detected with this new device. Now we are trying to develop a system which enables its quantitative measurement.(2) We examined the sphingolipid metabolism of peripheral blood cells, i.e., platelets, erythrocytes, neutrophils, and mononuclear cells. A distinguishing characteristic of sphingolipid metabolism in these highly-differentiated cells was their high sphingosine (Sph) kinase activity. About 40% of platelet Sph 1-phosphate (Sph-1-P) could be released extracellularly by 12-O-tetradecanoylphorbol 13-acetate, possibly through mediation by protein kinase C. On the other hand, in erythrocytes, neutrophils, and mononuclear cells, a significant percentage of Sph-1-P formed inside the cell was discharged without stimulation, while the stimulation-dependent release was marginal. Sph and its methylated derivative, N,N-dimethylsphingosine, induced apoptosis not only in neutrophils but also in mononuclear cells, while Sph-l-P elicited CaィイD22+ィエD2 mobilization in platelets. Our results suggest that all blood cells may remove plasma Sph, which is harmful or suppressive to cellular functions, and change it into Sph-1-P, acting as the source of plasma Sph-1-P, which may play a variety of important roles in blood vessels.