A versatile high-throughput assay to characterize antibody-mediated neutrophil phagocytosis
A versatile high-throughput assay to characterize antibody-mediated neutrophil phagocytosis
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DOI：
10.1016/j.jim.2019.05.006
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发表时间：
2019-08-01
影响因子：
2.2
通讯作者：
Alter, Galit
中科院分区：
文献类型：
--
作者：
Karsten, Christina B.;Mehta, Nickita;Alter, Galit
中性粒细胞是最丰富的白细胞，通过吞噬细胞清除、酶和免疫调节剂的分泌以及细胞外陷阱的释放，在抗病原体免疫中发挥着关键作用。中性粒细胞通过一系列先天免疫受体和炎症传感器非特异性地感知感染，但当抗体通过 Fc 受体利用时，也能够以病原体/抗原特异性方式做出反应。在中性粒细胞功能中，抗体依赖性中性粒细胞吞噬作用（ADNP）导致抗体介导的调理作用，使中性粒细胞能够以病原体适当的方式感知和响应感染。在这里，我们描述了一种高通量流式细胞术方法，可以有效地可视化和量化 ADNP 及其下游后果。该检测方法易于调整，支持纯化的中性粒细胞或白细胞的使用、纯化的 Ig 或血清的使用以及任何目标抗原的广泛用途。因此，该 ADNP 测定代表了深入表征中性粒细胞功能的高通量平台。
Neutrophils, the most abundant white blood cell, play a critical role in anti-pathogen immunity via phagocytic clearance, secretion of enzymes and immunomodulators, and the release of extracellular traps. Neutrophils nonspecifically sense infection through an array of innate immune receptors and inflammatory sensors, but are also able to respond in a pathogen/antigen-specific manner when leveraged by antibodies via Fc-receptors. Among neutrophil functions, antibody-dependent neutrophil phagocytosis (ADNP) results in antibody-mediated opsonization, enabling neutrophils to sense and respond to infection in a pathogen-appropriate manner. Here, we describe a high-throughput flow cytometric approach to effectively visualize and quantify ADNP and its downstream consequences. The assay is easily adaptable, supporting both the use of purified neutrophils or white blood cells, the use of purified Ig or serum, and the broad utility of any target antigen. Thus, this ADNP assay represents a high-throughput platform for the in-depth characterization of neutrophil function.