Tumor necrosis factor-α (TNF) and interferon-γ (IFN-γ) have been shown to regulate cell-mediated immunity and act as effective modifiers of immune function; however, their influence on neutrophil (PMN) function is not well defined. This study investigated the effect of these cytokines on PMN phagocytosis, respiratory burst, and complement receptor (C3b) expression. Human citrated whole blood was incubated with either phosphate-buffered saline (control), 20 μg Escherichia coli lipopolysaccharide (LPS), TNF (1, 10, or 100 units), or IFN-γ (1, 10, or 100 units). Synergy was also assessed between TNF and IFN-γ. Phagocytosis and respiratory burst were assayed by sequential incubation of blood with dichlorofluorescin diacetate followed by labeled Staphylococcus aureus. C3b receptor expression was assayed by labeled anti-CR3 monoclonal antibody. Measurements were expressed as the mean channel fluorescence of 2000 PMNs counted by flow cytometry. IFN-γ alone at all doses had no effect on any of the parameters measured. TNF 1 unit/ml increased phagocytosis (666 ± 47 vs 542 ± 19), respiratory burst (326 ± 33 vs 258 ± 17), and C3b (374 ± 42 vs 157 ± 14; all P < 0.05) over those of control. TNF also demonstrated dose-dependent PMN activation. The combination of TNF + IFN-γ increased both respiratory burst and C3b compared to either agent alone. These data indicate that TNF enhances PMN function and cytokine interaction may be important in PMN activation.