Figure 2
Effects of GKY25 on LPS responses of PMNs. PMN activation markers were assessed by flow cytometry on (A) isolated PMNS (5 × 105 cells/ml) co-treated with either 5 µM IVE25 or 5 µM GKY25 and 10 ng/ml LPS for 1 hr at 37 °C (n = 3) and (B) PMNs in whole blood co-treated with either 20 µM IVE25 or 20 µM GKY25 and 10 ng/ml LPS for 2 hrs at 37 °C (n = 3). PMNs stimulatory functions were assessed using the luminol-based ROS measurement – pre-warmed isolated PMNs (5 × 105 cells/ml) were co-treated with (C) either 5 µM IVE25 or 5 μM GKY25 and 10 ng/ml LPS (n = 5) and (D) 0.5, 1.0, 2.5, 5.0 or 10 µM GKY25 and 10 ng/ml LPS (n = 5). For positive control, PMNs in (C) were also treated with 25 nM PMA. (E) PMNs (5 × 105 cells/ml) pre-treated with 5 µM IVE25 (IVE25 > LPS) or 5 μM GKY25 (GKY25 > LPS) were also subjected ROS induction with 10 ng/ml LPS (n = 5). ROS measurements were done every 3 min for 3 hrs at 37 °C and AUCs were determined for statistical analyses. (−) Baseline. One-way ANOVA. Figures are representative of 3 independent experiments.
