Microfluidic arenas for war games between neutrophils and microbes
Microfluidic arenas for war games between neutrophils and microbes
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DOI：
10.1039/c8lc01263f
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发表时间：
2019-04-07
期刊：
影响因子：
6.1
通讯作者：
Irimia, Daniel
中科院分区：
文献类型：
--
作者：
Ellett, Felix;Jalali, Fatemeh;Irimia, Daniel
中性粒细胞活性（如细胞迁移和吞噬作用）的测量通常使用低含量批量测定法（其在单细胞水平上提供很少的详细活性）或流式细胞术方法（其具有单细胞分辨率但缺乏对过程动力学的看法）进行。在这里，我们提出了一种微流控检测方法，用于测量有助于中性粒细胞抗菌活性的基本功能：向靶标迁移和杀死微生物。该检测询问分离的人中性粒细胞和活的增殖微生物群体之间的相互作用。结果以反映体内感染的二元模式测量，体内感染被清除或耐受宿主反应。相互作用的结果也以中性粒细胞和微生物的单细胞分辨率表征。我们应用该测定来测试重症监护患者的中性粒细胞对活金黄色葡萄球菌的反应，并观察患者（包括败血症患者）的抗微生物中性粒细胞活性的改变。通过以高空间和时间分辨率直接测量中性粒细胞对活靶的活性，该测定提供了对塑造中性粒细胞和微生物群体之间相互作用结果的生死竞赛的独特见解。
Measurements of neutrophil activities such as cell migration and phagocytosis are generally performed using low-content bulk assays, which provide little detail activity at the single cell level, or flow cytometry methods, which have the single cell resolution but lack perspective on the kinetics of the process. Here, we present a microfluidic assay for measuring the essential functions that contribute to the antimicrobial activity of neutrophils: migration towards the target, and killing of microbes. The assay interrogates the interactions between isolated human neutrophils and populations of live, proliferating microbes. The outcome is measured in a binary mode that is reflective of in vivo infections, which are either cleared or endure the host response. The outcome of the interactions is also characterized at single cell resolution for both the neutrophils and the microbes. We applied the assay to test the response of neutrophils from intensive care patients to live Staphylococcus aureus, and observed alterations of antimicrobial neutrophil activity in patients, including those with sepsis. By directly measuring neutrophil activity against live targets at high spatial and temporal resolution, this assay provides unique insights into the life-or-death contest shaping the outcome of interactions between populations of neutrophils and microbes.