Figure 15.
Effect of LL-37 on the release of ectosomes from mouse bone marrow-derived neutrophils, and effect of ectosome administration on the survival of CLP mice. (a) Mouse bone marrow-derived neutrophils (2 × 106 cells) were stimulated with fMLF or LL-37 for 45 min at 37 °C, and the released ectosomes in the supernatant were analyzed by flow cytometry. Values were compared between the incubation with vehicle (PBS) and fMLF or LL-37, and among 1, 3 and 10 μg/mL of LL-37. An inlet shows a representative cytogram of ectosomes released from LL-37 (10 μg/mL)-stimulated neutrophils. (b) CLP mice were intraperitoneally injected with PBS (PBS-injected CLP) or ectosomes (3 × 105 ectosomes) isolated from LL-37-stimulated neutrophils (LL-37-MV-injected CLP) 2 h after the surgery. Sham mice were also injected with PBS (PBS-injected Sham) or ectosomes (LL-37-MV-injected Sham) 2 h after the surgery. The survival was monitored for 10 days, and the survival rate was compared between PBS-injected CLP and LL-37-MV-injected CLP mice by Kaplan-Meier method for 10 days and by chi square analysis for each day. * p < 0.05, ** p < 0.01,*** p < 0.001 [34].