Figure 1.
Regulation of TNF-α and IL-12 production by V-ATPase. Peritoneal macrophages (5 × 105) were cultured in 1 ml of normal medium for 2–3 hr, after which cells were treated for 9 or 24 hr with 0.1 μM bafilomycin A1, 10 μg/ml LPS, 100 units/ml IFN-γ, or combinations of reagents listed above. As bafilomycin was dissolved in DMSO, medium with an equal amount of DMSO (RPMI + Solvent) was used as control. TNF-α (A) was measured by bioassay and IL-12 (B) was determined by ELISA. Data represent means and standard deviations of duplicate cultures for each experimental condition, with TNF-α and IL-12 assays also performed in duplicate (n = 4). Similar results were obtained in four experiments with thioglycollate and in two experiments with resident peritoneal macrophages.