Fig. 5.
Impact of the preincubation for 1 h with LPS (100 ng/mL), LPS with LBP (2 ng/mL), fMLF (10 μM), and IL-8 (50 ng/mL), on I, F, and HBSS+/+ (as Ctrl) induced change in neutrophil cell shape and pHi. a In vitro effect of a preincubation with PAMPs or chemotactic agents on the neutrophil cell shape and on the ability of IL-8 and fMLF to alter the cell size. b pHi after in vitro exposure to the indicated molecules. The data are illustrated as merged violin plots with 2,000 neutrophils from independent donors to report precisely the distribution of the data. Discrepancies between individual donors did not explain the bimodal distribution of FSC and pHi on exposure to LPS. Preincubation with LPS reduced the IL-8-mediated response in FSC (c) and pHi (d). e Dose-effect response of the changes in FSC and pHi after stimulation with 0.1–1,000 ng/mL LPS and 2 ng/mL LBP for 1 h; n = 5–11. IL, interleukin; pHi, intracellular pH; HBSS+/+, Hank's Balanced Salt Solution; LBP, LPS-binding protein; PAMP, pathogen-associated molecular pattern; fMLF, N-formyl-Met-Leu-Phe; I, IL-8; F, fMLF; C, HBSS+/+; Ctrl, control; FSC, forward scatter area.