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首页> 外文期刊>Allergy >Establishment of a scalable microfluidic assay for characterization of population‐based neutrophil chemotaxis
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Establishment of a scalable microfluidic assay for characterization of population‐based neutrophil chemotaxis

机译：建立可扩展的微流体测定，用于表征群体中性粒细胞趋化性

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Abstract Background Regulation of neutrophil chemotaxis and activation plays crucial roles in immunity, and dysregulated neutrophil responses can lead to pathology as seen in neutrophilic asthma. Neutrophil recruitment is key for initiating immune defense and inflammation, and its modulation is a promising therapeutic target. Microfluidic technology is an attractive tool for characterization of neutrophil migration. Compared to transwell assays, microfluidic approaches could offer several advantages, including precis e control of defined chemokine gradients in space and time, automated quantitative analysis of chemotaxis, and high throughput. Methods We established a microfluidic device for fully automated, quantitative assessment of neutrophil chemotaxis. Freshly isolated mouse neutrophils from bone marrow or human neutrophils from peripheral blood were assessed in real time using an epifluorescence microscope for their migration toward the potent chemoattractants C‐X‐C‐motif ligand 2 (CXCL2) and CXCL8, without or with interleukin‐4 (IL‐4) pre‐incubation. Results Our microfluidic device allowed the precise and reproducible determination of the optimal CXCL2 and CXCL8 concentrations for mouse and human neutrophil chemotaxis, respectively. Furthermore, our microfluidic assay was able to measure the equilibrium and real‐time dynamic effects of specific modulators of neutrophil chemotaxis. We demonstrated this concept by showing that IL‐4 receptor signaling in mouse and human neutrophils inhibited their migration toward CXCL2 and CXCL8, respectively, and this inhibition was time‐dependent. Conclusion Collectively, our microfluidic device shows several advantages over traditional transwell migration assays and its design is amenable to future integration into multiplexed high‐throughput platforms for screening of molecules that modulate the chemotaxis of different immune cells.
机译：摘要中性粒细胞趋化性和激活的背景调节起到免疫力的关键作用，并且具有脱节的中性粒细胞反应可以导致中性嗜患者哮喘中所见的病理学。中性粒细胞招募是引发免疫防御和炎症的关键，其调节是有前途的治疗目标。微流体技术是一种有吸引力的介质迁移表征的有吸引力的工具。与Transwell测定相比，微流体方法可以提供若干优点，包括在空间和时间内定义的趋化因子梯度的Precis E控制，趋化性的自动定量分析和高通量。方法我们建立了一种微流体装置，用于全自动，定量评估中性粒细胞趋化性。从骨髓或人中性粒细胞的新鲜孤立的鼠嗜中性粒细胞使用eFforeence显微镜来评估从外周血的血液中的朝向有效的化学型C-X-C-MOTIF配体2（CXCL2）和CXCL8，没有或与白细胞介素-4迁移（IL-4）预孵育。结果我们的微流体装置允许精确和可再现的小鼠和人中性粒细胞趋化性的最佳CXCL2和CXCL8浓度的确定。此外，我们的微流体测定能够测量嗜中性粒细胞趋化性特异性调节剂的平衡和实时动态效应。我们通过表明小鼠和人性化粒细胞中的IL-4受体信号传导分别抑制它们对CXCL2和CXCL8的迁移，并且该抑制是时间依赖性的，通过抑制它们的迁移。结论集体，我们的微流体装置表现出传统的Transwell迁移测定方面的几个优点，其设计可用于将来集成到复用高通量平台中，以筛选调节不同免疫细胞的趋化性的分子。

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