Real-time deformability cytometry reveals sequential contraction and expansion during neutrophil priming.
Real-time deformability cytometry reveals sequential contraction and expansion during neutrophil priming.
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DOI：
10.1002/jlb.ma0718-295rr
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发表时间：
2019-06
影响因子：
5.5
通讯作者：
Toepfner N
中科院分区：
文献类型：
--
作者：
Bashant KR;Vassallo A;Herold C;Berner R;Menschner L;Subburayalu J;Kaplan MJ;Summers C;Guck J;Chilvers ER;Toepfner N
越来越明显的是，中性粒细胞的生物力学性质影响其通过循环，特别是通过肺毛细血管床的运输。最近提出，极化或形状改变的中性粒细胞在肺中的滞留有助于ARDS的发病机制。因此，本研究测试了中性粒细胞致敏和去致敏与能够改变细胞功能的形态流变学变化相关联的假设。我们采用实时变形性细胞术（RT-DC），这是一种最近开发的快速灵敏的方法，可以在几秒钟内评估数千个细胞的大小，形状和变形性的分布。在RT-DC分析过程中，由于中性粒细胞独特的粒度和大小，可以很容易地在抗凝的“全血”中识别出中性粒细胞，从而避免了对影响生物力学细胞特性的进一步分离技术的需要。因此，RT-DC是唯一适合描述形态流变学细胞变化的动力学。我们发现，激活或启动后，中性粒细胞经历了一个短时期的细胞收缩和硬化，随后细胞扩张和软化的阶段。在某些情况下，中性粒细胞最终恢复其未致敏的机械表型。人类中性粒细胞大小变化的潜在机制显示为Na+/H+反通道依赖性，并且预测其对中性粒细胞在健康和疾病中通过血管系统的运动具有深远意义。实时变形性细胞仪显示，致敏的中性粒细胞最初收缩，然后以Na+/H+反通道依赖性方式扩张，后一阶段与变形性增加相关
It has become increasingly apparent that the biomechanical properties of neutrophils impact on their trafficking through the circulation and in particularly through the pulmonary capillary bed. The retention of polarized or shape-changed neutrophils in the lungs was recently proposed to contribute to ARDS pathogenesis. Accordingly, this study tested the hypothesis that neutrophil priming and de-priming is coupled to morpho-rheological changes capable of altering cell function. We employ real-time deformability cytometry (RT-DC), a recently developed, rapid and sensitive way to assess the distribution of size, shape, and deformability of thousands of cells within seconds. During RT-DC analysis, neutrophils can be easily identified within anticoagulated ‘whole blood’ due to their unique granularity and size, thus avoiding the need for further isolation techniques, which affect biomechanical cell properties. Hence RT-DC is uniquely suited to describe the kinetics of morpho-rheological cell changes. We reveal that, following activation or priming, neutrophils undergo a short period of cell shrinking and stiffening, followed by a phase of cell expansion and softening. In some contexts, neutrophils ultimately recover their un-primed mechanical phenotype. The mechanism(s) underlying changes in human neutrophil size are shown to be Na+/H+ anti-port-dependent and are predicted to have profound implications for neutrophil movement through the vascular system in health and disease. Real-time deformability cytometry demonstrates that primed neutrophils initially contract, and then expand in a Na+/H+ anti-port-dependent manner, with the latter phase associated with increased deformability