Macrophage cathepsin B (CB) is implicated in tissue injury in inflammatory diseases. Lipopolysaccharide (LPS) is an activator of macrophages whose effect on CB is unknown. This study was undertaken to investigate the potential of macrophages as a source of increased CB and to determine if exposure to LPS might stimulate CB levels. As a model we chose the macrophage-like tumor line, THP-1. Incubation with LPS led to a time and dose-dependent increase in CB activity. LPS potentiated interferon-gamma (IFN-gamma)-induced elevations of CB and led to an additive increase in CB activity. Pretreatment of the cells with LPS not only caused a marked stimulation of CB activity over that seen with IFN-gamma alone, but also decreased the concentration and exposure time to the cytokine necessary to achieve maximum induction of the enzyme. The LPS and IFN-gamma induced CB increases were abolished by cycloheximide or actinomycin D in the cultures, indicating that the increases in CB required increased RNA transcription and de novo protein synthesis. Direct measurement of CB mRNA showed increases. These data indicate that although LPS alone appears to induce the production of CB in THP-1 cells, it augments IFN-gamma induced increases, suggesting that two signals are necessary for maximum CB induction in this system.