The past two decades have witnessed a resurgence in neutrophil research, inspired in part by the discovery of neutrophil extracellular traps (NETs) and their myriad roles in health and disease. Within the lung, dysregulation of neutrophils and NETosis have been linked to an array of diseases including pneumonia, cystic fibrosis, acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), and severe asthma. However, our understanding of pathologic neutrophil responses in the lung remains incomplete. Two methodologic issues have contributed to this gap: first, an emphasis on studying neutrophils from blood rather than the lung; and second, the technical difficulties of interrogating neutrophil responses in mice, which has largely restricted basic murine research to specialized laboratories. To address these limitations, we have developed a suite of techniques for studying neutrophil effector functions specifically in the mouse lung. These include ex vivo assays for phagocytosis and NETosis using bronchoalveolar neutrophils, and in situ evaluation of NETosis in a murine model of pneumonia. Throughout, we have prioritized technical ease and robust, quantitative readouts. We hope these assays will help to standardize research on lung neutrophils and improve accessibility to this burgeoning field.

中文翻译：

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小鼠肺炎中支气管肺泡中性粒细胞胞外陷阱和吞噬作用的定量。

过去二十年见证了中性粒细胞研究的复苏，部分原因是中性粒细胞胞外陷阱 (NETs) 的发现及其在健康和疾病中的无数作用。在肺内，中性粒细胞失调和 NETosis 与一系列疾病有关，包括肺炎、囊性纤维化、急性呼吸窘迫综合征 (ARDS)、慢性阻塞性肺病 (COPD) 和严重哮喘。然而，我们对肺部病理性中性粒细胞反应的理解仍然不完整。两个方法论问题导致了这一差距：首先，强调从血液而不是肺中研究中性粒细胞；其次，询问小鼠中性粒细胞反应的技术困难，这在很大程度上限制了专门实验室的基本鼠类研究。为了解决这些限制，我们开发了一套技术，专门用于研究小鼠肺中的中性粒细胞效应器功能。这些包括使用支气管肺泡中性粒细胞对吞噬作用和 NETosis 进行体外测定，以及在肺炎鼠模型中对 NETosis 进行原位评估。在整个过程中，我们优先考虑技术简易性和稳健的定量读数。我们希望这些检测有助于标准化肺中性粒细胞的研究，并提高对这一新兴领域的可及性。