Figure 3
IP-10 significantly enhances eosinophil O 2 - generation. (A) IP-10 significantly enhances eosinophil O2- generation in the presence of ICAM-1. Eosinophil cell density was adjusted to 1.25 × 106 cells/ml of HBSS/gel mixed 4:1 with cytochrome C, and the eosinophil suspension was then added to ICAM-1 coated 96-well plates. Immediately after adding IP-10 (30 or 100 nM), eosinophil O2- generation was measured based on the SOD-inhibitable reduction of cytochrome C. Data are shown as means ± SEM of 5 experiments using cells from different donors. (B) Eosinophil O2- generation induced by the combination of IP-10 + ICAM-1 is blocked by anti-β2 integrin mAb. Eosinophils were pre-incubated with anti-β2 integrin mAb (3 μg/ml) or control IgG for 15 min prior to analysis of O2- generation of eosinophils as in (A). Data are shown as means ± SEM of 7 experiments using cells from different donors. (C) Eosinophil O2- generation induced by a combination of IP-10 + ICAM-1 is blocked by anti-CXCR3 mAb. Eosinophils were pre-incubated with anti-CXCR3 mAb (3 μg/ml) or control IgG for 15 min prior to analysis of O2- generation of eosinophils as in (A). Data are shown as means ± SEM of 7 experiments using cells from different donors. Open and filled bars, eosinophil O2- generation without and with IP-10, respectively. *P < 0.05 versus without IP-10. #P < 0.05 versus mouse IgG1.