PT  - JOURNAL ARTICLE
AU  - Davide Scozzi
AU  - Carlos Puyo
AU  - Daniela Peruzzi
AU  - Kelsey Toth
AU  - Salvatore Mariotta
AU  - Andrew Gelman
TI  - LSC 2014 abstract - Endotracheal intubation triggers neutrophil extracellular traps
DP  - 2014 Sep 01
TA  - European Respiratory Journal
PG  - P3861
VI  - 44
IP  - Suppl 58
4099  - http://erj.ersjournals.com/content/44/Suppl_58/P3861.short
4100  - http://erj.ersjournals.com/content/44/Suppl_58/P3861.full
SO  - Eur Respir J2014 Sep 01; 44
AB  - Background: We have previously reported endotracheal intubation induces upper airway injury that leads to neutrophil recruitment in the absence of any evidence of infection. Neutrophil Extracellular Traps (NETs) are thought to play role in containing infection but it remains unclear if tissue injury stimulate NETs. Here we ask if endotracheal intubation can induce trachea-infiltrating neutrophils to produce NETs.Methods: Lavage above the tracheal cuff and peripheral blood was obtained from 3 healthy surgical patients that have undergone endotracheal intubation for general anesthesia. At the end of surgery tracheal lavage and peripheral blood neutrophils were cultured for 3 hrs with or without 25nM phorbol 12-myristate 13-acetate (PMA). NET formation was analyzed by fluorescence microscopy using antibodies specific for the NET components neutrophils elastase and histone protein H2b along with the DNA stain Hoescht 33342.Results: Both peripheral blood and tracheal lavage neutrophils produced NETs in response to PMA (Figure 1.A) However, unlike neutrophils in the peripheral blood tracheal lavage neutrophils spontaneously produced NETs in culture (Figure 1.B).Conclusions: Human infiltrating tracheal neutrophils in response to endotracheal intubation produce NETs. These data suggest that mechanically induced tissue injury is sufficient to promote NET formation.