Kupffer cells, in response to gut-derived bacteria, viruses and endogenous endotoxins, are thought to be a major source of proinflammatory cytokines and other mediators. In the present study, we investigated the mode of tumor necrosis factor alpha (TNF-alpha) and interleukin-1 (IL-1), interleukin-6 (IL-6) and prostaglandin E2 (PGE2) production by bovine Kupffer cells in response to LPS in vitro. By semi-quantitative reverse transcription-polymerase chain reaction, both TNF-alpha and IL-1beta mRNAs were expressed between 3 and 12 h after stimulation with 1 microg/ml of LPS. The IL-1alpha transcript also increased at 3 h, and then disappeared at 6 h. Although IL-6 mRNA was slightly expressed without stimulation, it increased and reached a peak after 6 h, and then decreased to normal levels by 24 h. The secretion of TNF-alpha and IL-1 was detected after 3 h and increased until 12 and 24 h, respectively, as detected by bioassay. IL-6 was secreted at a low level without stimulation, peaked 6 h after stimulation and remained elevated until 24 h. The secretion of prostaglandin E2 continued to increase for 24 h. These results suggest that several inflammatory cytokines released from bovine Kupffer cells are regulated in different modes.