Fig. 1: Rapid changes in PPP coupled to oxidative burst.
a, Heatmap showing metabolomic changes in human neutrophils activated ex vivo with zymosan (zym; 300 µg ml–1, 30 min), TNF-α (100 ng ml–1, 30 min), fMLP (100 nM, 10 min), PsA (10:1 PsA/neutrophils, 30 min) or PMA (100 nM, 30 min). Colour represents fold change compared to unstimulated neutrophils from the same donor. b, Oxidative burst induced by zymosan (500 µg ml–1), TNF-α (100 ng ml–1) or PMA (100 nM) (mean ± s.d., n = 6 technical replicates). c, NOX, stimulated by PMA and inhibited by DPI, produces superoxide using NADPH as the electron donor. OxPPP produces NADPH and can be inhibited by G6PDi or 6-AN. d,e, Oxidative burst induced by PMA (d) or zymosan (e) is inhibited by G6PDi, 6-AN or DPI (mean ± s.d., n = 6 (PMA), n = 8 (zymosan) technical replicates). f, Oxygen consumption by neutrophils incubated with PsA for 30 min with or without treatment by 6-AN, G6PDi or DPI (mean ± s.d., n = 4 technical replicates, indicated by dots). ND, not detectable (OCR for neutrophils not significantly different from zero). g, Heatmap showing metabolomic differences in human neutrophils stimulated ex vivo with zymosan (300 μg ml–1), PMA (100 nM), TNF-α (100 ng ml–1) or PsA for 30 min with or without DPI (10 µM) treatment. Colours indicate relative metabolite levels in conditions without DPI treatment compared to corresponding conditions (same donor, same stimulant) with DPI treatment. h, Changes in PPP and glycolytic intermediates over time upon stimulation with 100 nM PMA. Relative level is normalized to unstimulated neutrophils. *, Below reliable quantitation. Centre of the plot shows mean value from technical duplicates. i, Oxidative burst induced by varying PMA doses (mean ± s.d., n = 6 technical replicates). j, Relative abundances of PPP and glycolytic metabolites at 30 min after stimulation with varying doses of PMA. Bar graphs represent mean values of technical replicates (indicated by individual dots). b,d–f,h–j, Results confirmed in at least two independent experiments using neutrophils from different donors. a,g, Results from different donors presented in different rows. Ctrl, control.
