背景:中性粒细胞,人类最丰富的白细胞,在先天免疫中起关键作用,迅速迁移到感染和炎症的吞噬部位,中和,消灭入侵的病原体。中性粒细胞胞外捕获(NET)的形成越来越被认为是一种必不可少的快速先天免疫反应,但是当失调时,会导致败血症和免疫血栓形成疾病的发病机理。
目标:目前的NETosis模型有限,常规使用可以绕过天然NET调节途径的非生理触发因素。利用分离的中性粒细胞和永生化细胞系的模型,不反映中性粒细胞活化和NETosis的复杂生物学基础,发生在全血中。据我们所知,我们报道了第一个利用天然分子在全血中诱导NETosis的人离体模型。这种方法可用于药物筛选,重要的是,NETosis的无意活化剂。
方法:这里我们描述一个小说,在更生物学相关的Synthetic-Sepsis™模型中使用天然NETosis诱导因子的组合汇集的高通量离体全血诱导NETosis模型。
结果:我们发现不同的因素组合在NET产生率和/或NETosis大小方面引起不同的中性粒细胞反应。尽管供体之间存在差异,相似组的促炎分子在供体间诱导一致的应答.我们发现了至少三种生物触发因素,在我们的系统中诱导NETosis是必需的,包括TNF-α或LT-α。
结论:这些发现强调了在生物学相关背景下研究中性粒细胞生理学的重要性,以便更好地了解疾病病理学。危险因素,和治疗目标,潜在的,为疾病的干预和治疗提供新的策略。
BACKGROUND: Neutrophils, the most abundant white blood cells in humans, play pivotal roles in
innate immunity, rapidly migrating to sites of infection and inflammation to phagocytose, neutralize, and eliminate invading pathogens. Neutrophil Extracellular Trap (NET) formation is increasingly recognized as an essential rapid
innate immune response, but when dysregulated contributes to pathogenesis of sepsis and immunothrombotic disease.
OBJECTIVE: Current NETosis models are limited, routinely employing non-physiological triggers that can bypass natural NET regulatory pathways. Models utilizing isolated neutrophils and immortalized cell lines, do not reflect the complex biology underlying neutrophil activation and NETosis, that occurs in whole-blood. To our knowledge, we report the first human ex-vivo model utilizing naturally occurring molecules to induce NETosis in whole blood. This approach could be used for drug screening and, importantly, inadvertent activators of NETosis.
METHODS: Here we describe a novel, high-throughput ex-vivo whole blood induced NETosis model using combinatorial pooling of native NETosis inducing factors in a more biologically relevant Synthetic-Sepsis™ model.
RESULTS: We found different combinations of factors evoked distinct neutrophil responses in the rate of NET generation and/or magnitude of NETosis. Despite inter-donor variability, similar sets of pro-inflammatory molecules induced consistent responses across donors. We found at least three biological triggers, were necessary to induce NETosis in our system including either TNF-α or LT-α.
CONCLUSIONS: These findings emphasize the importance of investigating neutrophil physiology in a biologically relevant context to enable a better understanding of disease pathology, risk factors, and therapeutic targets, potentially, providing novel strategies for disease intervention and treatment.