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. Overall design: 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.