Figure 2.

Contact of swine trachea with an ETT promotes neutrophil migration, necrosis, and an activated neutrophil phenotype during presence of mtDNA. 2A) Total live neutrophils cell count was done by tryptan blue exclusion following negative immunomagnetic isolation in coated (n = 5, gray) and uncoated ETT (n = 5, black) groups over a period of 6 h. A high neutrophil count was observed in the uncoated group. 2B) Cellular viability analysis was conducted in SSC^lo‐hi for AnnexinV^lo‐hi and FSC for 7AAD^lo‐hi. Left FACS panel shows elevated necrosis (9.70%) and low early apoptosis (2.65%), high late apoptosis (17.2%) with live cells accounting for (70.5%) in the untreated ETT group, whereas, the right panel shows lower necrosis and late apoptosis and higher concentration of live cells in the coated group. Statistical analysis is reported in the supplemental data Fig. 2B. 2C) Tracheal lavage fluid (TLF) analysis was conducted by flow cytometry in coated (gray bars n = 5) and uncoated ETT (black bars n = 5), with neutrophil cells identified by CD16^lo‐hi/CD62L^lo‐hi monoclonal antibodies. Neutrophils with the phenotype (CD16^lo/CD62L^hi/CD11a^hi/CD11b^hi/CD18^hi/CD54^hi) are indicative of activation and migration as determined in TLF. Neutrophils of uncoated samples showed a higher surface expression of ICAM‐1 and integrins as compared to the uncoated group. Data shown represents MFI ± sem *P < 0.05; **P < 0.01, derived from 10 swine experiments