Fig. 1: ARDS induces long-term alterations of neutrophil phenotypes and functions.
a–i, Phenotypic and functional analysis of circulating neutrophils collected from healthy controls and survivors of ARDS 3–6 months post-hospital admission. a, Schematic representation of the analyses performed. Human silhouette adapted from Wikipedia commons (https://commons.wikimedia.org/wiki/File:Man_shadow_-_upper.png); lungs, neutrophils and blood tube adapted from Servier license under CC-BY 3.0 Unported. b, Circulating neutrophil counts obtained by flow cytometry (n = 19 healthy control and n = 26 ARDS survivor). c,d, Surface expression of the neutrophil activation markers CD66b and CD62L measured by flow cytometry (n = 15 healthy control and n = 14 ARDS survivor (c), n = 10 healthy control and n = 13 ARDS survivor (d)). e, Metabolite abundance of pyruvate, lactate and acetyl-CoA obtained by LC–MS analysis (n = 5 for both experimental groups). f, Proteomic analysis by LC–MS showing abundance of cytoskeletal (GO:0005856) and azurophilic granule cargo proteins (GO:0035578) (n = 8 for both experimental groups). g, Ex vivo quantification of α-1-antitrypsin from neutrophil culture supernatants performed by ELISA (n = 4 for both experimental groups, two technical replicates per sample). h, Ex vivo neutrophil survival in response to LPS stimulation after 20 h of culture evaluated by microscopy analysis (n = 7 for both experimental groups, two technical replicates per sample). i, Phagocytic capacity of opsonized S. aureus SH1000 measured by flow cytometry as a percentage of the total neutrophil population analyzed (n = 4 for both experimental groups, two technical replicates per sample). j, Infections recorded in ARDS survivors, with their infection etiologies expressed as proportion or as cumulative positive microbiology results over the course of 6 months post-ARDS. b–i, Data show mean ± s.d., with each value representing an individual. Significant P values depicted (for P < 0.05) and obtained by Shapiro–Wilk normality test followed by a two-tailed t-test (b–i) or two-tailed Mann–Whitney U-test (c, cytoskeleton (f)). FC, fold change; gMFI, geometric mean fluorescence intensity; a.u., arbitrary units.
