Figure 2
Chemokine secretion by stimulated primary small airway epithelial cells (SAEC). (A) SAEC were stimulated for 24 h with different TLR ligands including poly(I:C), flagellin, zymosan and macrophage activating lipopetide-2 (MALP-2) and cell supernatants were analyzed using multiplex ELISAs. Results are shown as fold changes relative to untreated controls. The figure shows representative results of three independent experiments. Stimulation of the cells with LPS or PGN had no significant effect on the secretion of the indicated chemokines (data not shown). (B) Inhibition of poly(I:C) induced IP-10 secretion by a monoclonal, functional blocking anti-TLR3 antibody. SAEC were stimulated in the presence of an anti-TLR3 antibody or an IgG1 isotype control with 5 μg/ml poly(I:C) for 6 h and IP-10 secretion was analyzed in triplicates using an IP-10 ELISA. Results were compared to untreated controls and poly(I:C) stimulated cells in the absence of an antibody. (C) IFN-β secretion of SAEC after stimulation with poly(I:C). SAEC were stimulated with 5 μg/ml poly(I:C) for 6 h or 24 h and IFN-β secretion was analyzed in triplicates using an IFN-β ELISA. Results were compared to untreated controls. (D) Inhibition of poly(I:C) induced IP-10 and I-TAC expression by a goat polyclonal functional blocking anti-IFN-β antibody. SAEC were stimulated in the presence of a goat anti-IFN-β antibody (1 or 5 μg/ml; gray bars) or a goat IgG control (1 or 5 μg/ml; white bars) with 5 μg/ml poly(I:C) for 6 h. IP-10 or I-TAC expression was analyzed by real-time RT-PCR. Expression data were normalized using β-actin as endogenous control and are shown as fold changes relative to untreated controls. Results were compared to poly(I:C) stimulated cells in the absence of an antibody (black bars).