Figure 5
Effect of anti-TLR4 blocking antibody on endotoxin-induced superoxide production by PBMC. Human PBMC (~1.0 × 106 total cells/well, 100,000 monocytes) were prepared in 100 μl of incubation buffer containing 2 μg/well of mouse non-immune IgG or of HTA-125, and were placed in individual wells of a 96-well white Optiplate. Plates were incubated at 37°C for 30 min. Plates were then cooled on ice, and 100 μl of incubation buffer containing 0.2 mM lucigenin and 2× the final indicated concentration of endotoxin was added to the wells. The plates were then placed in the luminometer and two-sec readings were taken at 37°C every 2 min for 180 min. The resulting readings in relative light units (LU) were summed to provide a measure of activity. Data are expressed as total LU (in thousands) per 100,000 monocytes and represent the mean of duplicate samples. There were no significant differences between the mouse IgG concentration curve and the HTA-125 concentration curve.