BACKGROUND The hallmark of the cystic fibrosis (CF) lung disease is a neutrophil dominated lung environment that is associated to chronic lung tissue destruction and ultimately the patient's death. It is unclear whether the exacerbated neutrophil response is primary related to a defective CFTR or rather secondary to chronic bacterial colonization and inflammation. Here, we hypothesized that CF peripheral blood neutrophils present intrinsic alteration at birth before the start of an inflammatory process. METHODS Peripheral blood neutrophils were isolated from newborn CFTR+/+ and CFTR-/- piglets. Neutrophils immunophenotype was evaluated by flow cytometry. Lipidomic and proteomic profile were characterized by liquid chromatography/tandem mass spectrometry (LC-MS/MS), intact cell matrix-assisted laser desorption/ionization mass spectrometry (ICM-MS) followed by top-down high-resolution mass spectrometry (HRMS), respectively. The ability of CF neutrophils to kill pseudomonas aeruginosa was also evaluated. RESULTS Polyunsaturated fatty acid metabolites analysis did not show any difference between CFTR+/+ and CFTR-/- neutrophils. On the other hand, a predictive mathematical model based on the ICM-MS proteomic profile was able to discriminate between both genotypes. Top-down proteomic analysis identified 19 m/z differentially abundant masses that corresponded mainly to proteins related to the antimicrobial response and the generation of reactive oxygen species (ROS). However, no alteration in the ability of CFTR-/- neutrophils to kill pseudomonas aeruginosa in vitro was observed. CONCLUSIONS ICM-MS demonstrated that CFTR-/- neutrophils present intrinsic alterations already at birth, before the presence of any infection or inflammation.

中文翻译：

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囊性纤维化新生仔猪外周中性粒细胞的内在改变

背景囊性纤维化(CF)肺病的标志是中性粒细胞占主导的肺环境，其与慢性肺组织破坏和最终患者的死亡相关。目前尚不清楚加剧的中性粒细胞反应是否主要与有缺陷的 CFTR 相关，还是继发于慢性细菌定植和炎症。在这里，我们假设 CF 外周血中性粒细胞在炎症过程开始之前在出生时呈现内在改变。方法 从新生 CFTR+/+ 和 CFTR-/- 仔猪中分离外周血中性粒细胞。通过流式细胞术评估嗜中性粒细胞免疫表型。脂质组学和蛋白质组学特征通过液相色谱/串联质谱法 (LC-MS/MS) 进行表征，完整的细胞基质辅助激光解吸/电离质谱 (ICM-MS)，然后分别是自上而下的高分辨率质谱 (HRMS)。还评估了 CF 嗜中性粒细胞杀死铜绿假单胞菌的能力。结果 多不饱和脂肪酸代谢物分析未显示 CFTR+/+ 和 CFTR-/- 中性粒细胞之间的任何差异。另一方面，基于 ICM-MS 蛋白质组学特征的预测数学模型能够区分两种基因型。自上而下的蛋白质组学分析确定了 19 m/z 差异丰富的质量，主要对应于与抗菌反应和活性氧 (ROS) 产生相关的蛋白质。然而，没有观察到 CFTR-/- 中性粒细胞在体外杀死铜绿假单胞菌的能力发生变化。