Saliva-induced NETosis is independent on NADPH oxidase or elastase and dependent on the MEK/ERK pathway. NETosis was stimulated in neutrophils and examined by immunofluorescence microscopy. NET formation was quantified as described in “Materials and methods.” Representative images are shown in supplementary Figure 3. (A) Neutrophils from 2 patients with chronic granulomatous disease (CGD) were stimulated with PMA or glucose oxidase (GO) for 2.5 hours or with saliva for 1 hour. Results are shown as the average from the 2 CGD patients. (B) Neutrophils from patient with PLS were stimulated for 1 hour with saliva. Results are shown as the average of 2 independent experiments with neutrophils PLS patient. (C) Neutrophils were preincubated with inhibitors or solvent (DMSO) before stimulation with saliva in the presence of inhibitors. Results are shown from 3 independent experiments. Scale bars, 100 μm. (D) Binding of various proteins to NETs isolated from morning saliva or induced with saliva, bacteria, or PMA. Control depicts nonstimulated neutrophils not undergoing NETosis. Columns denote average values and error bars standard deviations. Scale bars, 10 μm. *P < .05; **P < .01 and refer to nominal significance of inhibition or induction of NET formation.