Neonatal monocytes in tracheal aspirates from infants at risk for Bronchopulmonary Dysplasia propagate IL-1 signaling in the first weeks of lung injury

Sorted human monocytes, isolated from tracheal aspirates from intubated premature infants. Overall design: We prospectively recruited premature infants who were born at less than 29 weeks gestation and who were intubated/ventilated for this study. Tracheal aspirate samples were obtained at several time points (d0-3, 7-9, 14-17, 28-31) when possible. Samples were delivered to BRI for analysis. We used a 13-color panel of conjugated antibodies to identify T cells, B cells, NK cells, basophils, neutrophils, eosinophils, and 2 populations of monocyte/macrophages CD14+CD16+ (DP) and CD14 CD16- (SP) by flow cytometry. Samples were sorted on the Aria II, and DP and SP cells were sorted into lysis buffer (Clontech/Takara RNA isolation kit). Total RNA from these samples was isolated by us and delivered to the genomics core for cDNA synthesis with Smartseq