Figure 7
ES-62 modulates IL-33 responses. PDMCs were sensitized with murine anti-DNP IgE (0.5 μg/ml) in the presence or absence of ES-62 (2 µg/ml) overnight and then incubated in medium ± IL-33 (10 ng/ml) for 24 h at 37 °C before release of IL-6, IL-13 and CCL2 was measured by ELISA (a–c). Data are mean ± SD values where ***p < 0.001 and are from single experiments, which for IL-6 and IL-13 were representative of four-six experiments. Analysis of the latter data shows that ES-62 significantly reduces IL-33-induced IL-6 and IL-13 production by PDMCs to 66.03 ± 16.82 n = 5, p < 0.05 and 59.85 ± 14 n = 6, p < 0.01 of the control responses respectively and by BMMCs to 81.23 ± 5.72 n = 4, p < 0.05 and 75.75 ± 8.13 n = 5, p < 0.05 of the control responses respectively. PDMCs (d) were sensitised with murine anti-DNP IgE (0.5 μg/ml) in the presence or absence of ES-62 (2 µg/ml) and then loaded with Fura-2/AM and stimulated at 50 s in serum-free HBSS with IL-33 (10 ng/ml). Calcium mobilisation data are presented as the mean (baseline subtracted) values ± SEM where for IL-33, n = 3 and for IL-33 + ES-62, n = 4 replicate biological samples from a single experiment representative of at least three independent experiments. Statistical analysis was by two-way ANOVA where *** p < 0.001. WT and ST2−/− PDMCs (e,f) and BMMCs (g,h) were sensitized with murine anti-DNP IgE (0.5 μg/ml) in the presence or absence of ES-62 (2 µg/ml) overnight and then incubated in medium ± 0.5 µg/ml DNP-HSA plus 10 ng/ml IL-33 (XL + IL-33) for 24 h at 37 °C before release of IL-6 (e,g) and IL-13 (f,h) was measured by ELISA. Data are mean ± SD values where ***p < 0.001. Analysis of 3 independent experiments showed that ES-62 reduced IL-33 + XL-stimulated IL-6 and IL-13 production by PDMCs to 79.27 ± 12.42 and 59.98 ± 10.07, p < 0.05 of the control responses respectively and by BMMCs to 65.11 ± 26.72 and 65.60 ± 20.43, p < 0.05 of the control responses in WT, but not ST2KO mast cells.
