An Experimentally Determined State Diagram for Human CD4+ T Lymphocyte CXCR4-Stimulated Adhesion Under Shear Flow.
An Experimentally Determined State Diagram for Human CD4+ T Lymphocyte CXCR4-Stimulated Adhesion Under Shear Flow.
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实验确定的剪切流下人 CD4 T 淋巴细胞 CXCR4 刺激粘附的状态图。
DOI：
10.1007/s12195-018-0519-x
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发表时间：
2018
影响因子：
2.8
通讯作者：
Hammer,DanielA
中科院分区：
文献类型：
--
作者：
Anderson,NicholasR;Lee,Dooyoung;Hammer,DanielA
简介白细胞粘附级联对于维持体内平衡以及免疫细胞进入感染和炎症部位的能力非常重要。尽管进行了大量工作来识别级联的分子成分，并进行了大量模拟来预测分子密度、身份和粘附之间的关系，但这些关系尚未通过实验测量。方法使用重组 ICAM-1 和/或 E-选择素以及固定化 SDF-1α 功能化的表面，我们使用流动室来测量原代幼稚人 CD4+T 淋巴细胞在不同表面密度的配体上的束缚、滚动和停滞速率。结果细胞需要最低水平的配体密度超越束缚。研究发现 E-选择素和 ICAM-1 在促进细胞停滞方面具有协同关系。具有两种配体的表面具有最高水平的阻滞，而仅含有 E-选择素的表面阻碍了细胞超越滚动的能力。相比之下，ICAM-1 的表面只允许束缚或逮捕。细胞保持恒定的滚动速度和时间，以阻止表面密度和成分的巨大变化。此外，只有O(101)个位点/μm2的表面密度允许滚动，而O(102)个位点/μm2的表面密度促进阻滞，大约等于先前确定的模拟值。结论我们系统地和实验性地绘制了流动下T细胞粘附的状态图，直接展示了每种动态粘附状态的定量要求，并展示了多个粘附分子如何协同作用以确保阻滞。
IntroductionThe leukocyte adhesion cascade is important for the maintenance of homeostasis and the ability of immune cells to access sites of infection and inflammation. Despite much work identifying the molecular components of the cascade, and numerous simulations to predict the relationship between molecule density, identity, and adhesion, these relationships have not been measured experimentally.MethodsUsing surfaces functionalized with recombinant ICAM-1 and/or E-selectin along with immobilized SDF-1α, we used a flow chamber to measure rates of tethering, rolling and arrest of primary naïve human CD4+T lymphocytes on different surface densities of ligand.ResultsCells required a minimum level of ligand density to progress beyond tethering. E-selectin and ICAM-1 were found to have a synergistic relationship in promoting cell arrest. Surfaces with both ligands had the highest levels of arrest, while surfaces containing only E-selectin hindered the cell’s ability to progress beyond rolling. In contrast, surfaces of ICAM-1 allowed only tethering or arrest. Cells maintained constant rolling velocity and time to stop over large variations in surface density and composition. In addition, surface densities of only O(101) sites/µm2allowed for rolling while surface densities of O(102) sites/µm2promoted arrest, approximately equal to previously determined simulated values.ConclusionsWe have systematically and experimentally mapped out the state diagram of T cell adhesion under flow, directly demonstrating the quantitative requirements for each dynamic state of adhesion, and showing how multiple adhesion molecules can act in synergy to secure arrest.