Kinetics of the different steps during neutrophil migration through cultured endothelial monolayers treated with tumour necrosis factor-α
Kinetics of the different steps during neutrophil migration through cultured endothelial monolayers treated with tumour necrosis factor-α
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DOI：
10.1159/000025690
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发表时间：
1999-11-01
影响因子：
1.7
通讯作者：
Nash, GB
中科院分区：
文献类型：
--
作者：
Luu, NT;Rainger, GE;Nash, GB
为了更好地了解中性粒细胞迁移的调节，我们研究了内皮细胞单层上、下和通过内皮细胞单层的粘附和迁移动力学。将中性粒细胞灌注在已用肿瘤坏死因子-α（TNF; 2- 1，000 U/ml）处理4小时的人脐静脉内皮细胞（HUVEC）上。视频显微镜检查显示，经内皮迁移完成约5分钟内完成的灌注团的中性粒细胞。然后可以观察到粘附细胞的单独群体，以不同的特征速度在HUVEC表面上滚动、迁移或在下方迁移。增加TNF浓度对迁移动力学影响不大，但将平衡从滚动粘附转移到跨内皮迁移。当从单个中性粒细胞与用100 U/ml TNF处理的HUVEC结合的那一刻起对其进行跟踪时，我们发现，类似于40%的中性粒细胞在接触时基本上立即固定，而类似于40%的中性粒细胞在滚动不同的时间段（平均26秒）后固定，并且类似于20%的中性粒细胞连续滚动。大多数固定的细胞在花费20-200秒迁移到顶部后继续迁移通过单层，并且花费约60秒通过。总体而言，从首次结合到完成迁移的时间平均为152 s（范围类似于60-240 s）。有趣的是，中性粒细胞在单层的顶部移动相对较慢（约8 μ m/min），但在下面移动更快（约16 μ m/min）。我们认为，中性粒细胞迁移过程中的不同阶段具有不同的控制机制的特征动力学，这对血管系统的清除效率和速率产生了重要影响。版权所有（C）1999 S. Karger AG，巴塞尔。
To enable a better understanding of the regulation of neutrophil migration, we investigated the kinetics of adhesion and migration over, through and under endothelial monolayers. Neutrophils were perfused over human umbilical vein endothelial cells (HUVEC) which had been treated with tumour necrosis factor-alpha (TNF; 2-1,000 U/ml) for 4 h. Videomicroscopy showed that transendothelial migration was complete within about 5 min of completion of perfusion of a bolus of neutrophils. Separate populations of adherent cells could then be observed, either rolling, migrating over the surface of the HUVEC or migrating underneath, at different characteristic speeds. Increasing concentration of TNF had little effect on the kinetics of migration, but shifted the balance from rolling adhesion to transendothelial migration. When individual neutrophils were followed from the moment they bound to HUVEC treated with 100 U/ml TNF, we found that similar to 40% immobilised essentially immediately on contact, while similar to 40% immobilised after rolling for varying periods (average 26 s) and similar to 20% rolled continuously. Most of the immobilised cells went on to migrate th rough the monolayer after spending 20-200 s migrating on top, and took about 60 s to pass through. Overall, the time from first binding to completion of transmigration averaged 152 s (range similar to 60-240 s). Interestingly, neutrophils moved relatively slowly on top of the monolayer (about 8 mu m/min) but more rapidly underneath (about 16 mu m/min). We suggest that the different stages during neutrophil transmigration have characteristic kinetics with separate control mechanisms, which critically influence the efficiency and rate of clearance from the vasculature. Copyright(C) 1999 S. Karger AG, Basel.