Effects of AT on neutrophil extracellular ROS production and microcapillary passage. Purified human neutrophils were incubated with different concentrations of HSA, HRG, AT, rAT, ARG, or HBSS at 37°C. Extracellular ROS production of neutrophils was determined using isoluminol and HRP. (A) Time-course changes in neutrophil extracellular ROS production after addition of each reagent. (B) The bar graphs show the neutrophil extracellular ROS production at 30 minutes under different conditions. ∗∗∗P < .001 vs HBSS, †††P < .001 vs HSA. (C) The concentration-response curves for the effects of AT, rAT, and ARG on the neutrophil extracellular ROS production assay. The purified human neutrophils were incubated with HSA, AT, or HBSS at 37°C for 60 minutes and then applied to a microchannel flow analyzer (MC-FAN). (D) The results showed an image of the microchannel at the passage of 100 μL. (E) The adhesive neutrophils that stayed inside the capillaries after 100-μL passage were counted. The results are the means ± SE of 3 wells for panels A-C or 3 determinations for panel E. RLU, relative light unit.