A, Eosinophil surface expression of CD69 after overnight incubation in culture medium (CM) +/− GM-CSF (50ng/mL). The antibodies were crosslinked with F(ab’)2 goat anti-mouse (10 μg/mL). Measurement of EPX (B), TNFα (C) and IL-8 (D) in culture supernatants following crosslinking of isotype control (white bars), anti-Siglec-7 (dark gray bars), or anti-Siglec-8 (light gray bars) at antibody concentrations ranging from 0.62–2.5 μg/mL on eosinophils for 40 minute (EPX), or overnight (TNFα and IL-8) in the presence of GM-CSF (n=7–8). Mediator release is expressed as the % of the level measured in the control well containing 50 ng/mL GM-CSF alone. Data are expressed as % GM with 95% CI. *P < .05; **P < .001; Wilcoxon matched-pairs signed rank test. Symbols represent individual subjects (white = ND).