Although ROS can participate in modulating the activity of the transcriptional factor NF-kappaB and expression of NF-kappaB-dependent genes, the mechanisms involved and the roles of specific ROS have not been fully determined. In particular, individual ROS appear to have differing effects on NF-kappaB activation dependent on the cell population studied. In the present study, we examined the ability of H(2)O(2) to affect NF-kappaB activation in LPS-stimulated murine neutrophils and macrophages. Exposure of bone marrow or peritoneal neutrophils to H(2)O(2) was associated with reduced nuclear translocation of NF-kappaB and decreased production of the NF-kappaB-dependent cytokines TNF-alpha and macrophage inhibitory protein-2. H(2)O(2) treatment resulted in diminished trypsin- and chymotrypsin-like proteasome activity. The degradation of IkappaB-alpha normally found in LPS-treated neutrophils was prevented when H(2)O(2) was added to cell cultures. In contrast to the effects found in neutrophils, H(2)O(2) did not affect chymotrypsin-like proteasomal activity or cytokine production in LPS-stimulated macrophages, even though trypsin-like proteasomal activity was reduced. These results demonstrate that the effects of H(2)O(2) on NF-kappaB and proteasomal activity are cell population specific.