The effect of norepinephrine (NE) on the production of tumor necrosis factor (TNF) by rat spleen macrophages was determined. Following activation with lipopolysaccharide, analysis of both secreted and cell-associated samples showed that TNF activity was significantly suppressed in the presence of 10 microM NE. With the addition of the beta-receptor antagonist propranolol a partial reversal of the suppressive effect of NE was noted whereas the addition of the mixed alpha-receptor antagonist phentolamine induced a more pronounced suppressive effect in the supernatant fraction. Similar results were obtained with epinephrine and isoproterenol. Control experiments confirmed that this effect of NE was mediated at the level of macrophage activation. Analysis of lymphocyte activating factors demonstrated a similar pattern of response. Since factors released by macrophages participate in many aspects of the immune response, these results support a functional role for sympathetic innervation of the spleen in immune regulation.