Abnormal Immune Programming in Neutrophils of Cystic Fibrosis

This study performed single-cell RNA sequencing (scRNA-seq) analyses on peripheral blood neutrophils from cystic fibrosis (CF) patients and healthy controls (HC) in their native state and LPS-activated state. Differences in global gene transcription were interrogated and compared. Results demonstrate that CFTR loss of function in CF neutrophils led to abnormal immune programming, exemplified by precocious priming at their native state, and abnormal immune response to LPS. Peripheral blood (~10 cc) from cystic fibrosis patients and healthy controls was drawn using a BD EDTA-purple top Vacutainer tube. Neutrophils were isolated by Percoll-density gradient centrifugation method, and were split into two aliquots. One was immediately processed for sc-RNA-seq, and the other was cultured and stimulated with Pseudomonas aeruginosa LPS (40 µg/ml) for 16 hours, followed by scRNA-seq.

本研究针对囊性纤维化（cystic fibrosis, CF）患者与健康对照（healthy controls, HC）的外周血中性粒细胞，分别在天然状态与脂多糖（lipopolysaccharide, LPS）激活状态下开展了单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）分析，对全局基因转录水平的差异进行了探究与对比；研究结果显示，CF患者中性粒细胞内CFTR功能缺失可引发异常免疫编程，具体表现为天然状态下的早熟致敏，以及对LPS的异常免疫应答。研究人员使用BD EDTA紫色帽Vacutainer真空采血管，采集囊性纤维化患者与健康对照的外周血样本（约10 cc），通过Percoll密度梯度离心法分离得到中性粒细胞后，将其分为两份：一份即刻进行scRNA-seq测序，另一份进行体外培养，以铜绿假单胞菌（Pseudomonas aeruginosa）来源的LPS（40 µg/ml）刺激16小时后，再开展scRNA-seq测序。