Neutrophil extracellular traps released in response of cigarette smoke extract increases the secretion of proinflammatory cytokines by human alveolar epithelial cells

SABBIONE, F; GALLETTI, J; TOWSTYKA, N; KEITELMAN, I; FERRERO, MC; JANCIC, C; GUZMÁN, M; BALDI, PC; TREVANI, A

Mar del Plata

Congreso; LXIV Reunión Anual de la Sociedad Argentina de Inmunología; 2016

Sociedad Argentina de Inmunología

Cigarette smoking is a major causeof chronic obstructive pulmonary disease. Gas phase of cigarette smoke (CS) canreach the alveolar epithelium inducing the secretion ofchemokines and cytokinesthat contribute to recruit neutrophils (PMN) into the airway lumen. In responseto diverse stimuli,PMNreleaseneutrophil extracellular traps (NET) through a processcalled NETosis.Previous studies determined thatCS and uric acid (UA),a majorDAMP found at high concentrations in lungsof patients with acutelung injury,can induce NETosis.The aim of this study was to determine if a short exposureto CS is able to induce NETosis, and if theseNET induce proinflammatorycytokine secretion by alveolar epithelial cells,comparing their effects withthoseproduced by NET released in response to UA(8 mg/dl).CSextract (CSE)comingfrom cigarettes containing an equivalent of 13.6 mg/ml tar prepared accordingto a standard methodwas used at 10%. PMN were incubated with or withoutstimulifor 1h, then washed and cultured for 3 more hours at 37°C. NET were identified byconfocal microscopy by colocalization of DNA with myeloperoxidase. Shortexposure to both stimuli induced NETosis. NET released by 106 PMN wereisolated and added to A549 epithelial cell monolayers.We found that bothCSE-andUA-induced NET significantly increased the secretion of IL-8 andIL-6(p