Activation of integrin αIIbβ3 expressed in Chinese hamster ovary cells is required for interaction with solid-phase von Willebrand factor
Activation of integrin αIIbβ3 expressed in Chinese hamster ovary cells is required for interaction with solid-phase von Willebrand factor
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DOI：
10.1046/j.1365-2141.2002.03960.x
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发表时间：
2002-12-01
影响因子：
6.5
通讯作者：
Baruch, D
中科院分区：
文献类型：
--
作者：
Mekrache, M;Legendre, P;Baruch, D
已知二硫苏糖醇（DTT）可诱导α IIb β 3整联蛋白的活性构象，并在可溶性纤维蛋白原（Fg）存在下促进中国仓鼠卵巢（CHO）-α IIb β 3细胞聚集。本研究的目的是比较在存在或不存在DTT的情况下，CHO-α IIb β 3细胞的Fg或血管性血友病因子（VWF）的粘附和扩散。我们的研究结果表明，DTT处理是需要诱导细胞在VWF上铺展。相比之下，在没有DTT的情况下，CHO-alphaIIbbeta 3细胞在Fg上的扩散已经是最佳的。我们使用了一个小的灌注室耦合到视频显微镜，以证明CHO-alphaIIbbeta 3细胞的粘附和蔓延VWF需要DTT活化，以抵抗在增加剪切速率（50-1600/s）下的分离。相比之下，未处理的或经DTT处理的细胞在Fg上铺展能够抵抗极高的流速。这些数据提供了新的证据，活化的alphaIIbbeta 3是绝对需要的扩散细胞抵抗脱离，并加强了粘附和扩散到VWF的alphaIIbbeta 3活化步骤的重要性。
Dithiothreitol (DTT) is known to induce an active conformation of alphaIIbbeta3 integrin and to promote the aggregation of Chinese hamster ovary (CHO)-alphaIIbbeta3 cells in the presence of soluble fibrinogen (Fg). The aim of this study was to compare adhesion and spreading with Fg or von Willebrand factor (VWF) of CHO-alphaIIbbeta3 cells in the presence or absence of DTT. Our results indicate that DTT treatment was required to induce cell spreading on VWF. In contrast, CHO-alphaIIbbeta3 cell spreading on Fg was already optimal in the absence of DTT. We used a small perfusion chamber coupled to videomicroscopy to demonstrate that CHO-alphaIIbbeta3 cells that were adherent and spread on VWF required DTT activation to resist to detachment under increasing shear rates (50-1600/s). In contrast, untreated or DTT-treated cells spread on Fg were able to resist to extremely high flow rates. These data provide novel evidence that activated alphaIIbbeta3 is absolutely required for spread cells to resist detachment and strengthens the importance of the alphaIIbbeta3 activation step for adhesion and spreading to VWF.