To evaluate the role of the dialysate in the stimulation of interleukin-1 (IL-1) production during clinical hemodialysis (HD), we studied maintenance HD patients in two experiments. Cellulosic hollow-fiber dialyzers were obtained after 20 minutes of HD using either nonsterile standard dialysate (n = 6) or sterile pyrogen free 0.9% saline as dialysate (n = 6). After rinsing the blood compartment with normal saline, dialyzers were incubated at 37 degrees C for six hours. Aliquots from the blood compartment were analyzed for the presence of IL-1 by (1) rabbit pyrogenic response after intravenous injection or (2) thymocyte co-proliferation assay. The in vivo assay showed a significantly greater febrile response when standard dialysate was used than in the sterile saline group (P less than .001), and this response could be abolished by heat inactivation of aliquots (P less than .001). The in vitro assay confirmed the presence of significantly greater amounts of IL-1 (P less than .05). Studies were repeated using filter sterilized standard dialysate (n = 6) v standard dialysate (n = 6) for 240 minutes of clinical HD. The in vitro assay revealed significantly lower IL-1 levels in the filtered sterilized dialysate group (P less than .05), however, a blank control assay showed yet significantly lower levels (P less than .05). We conclude that IL-1 is produced during clinical HD and that endotoxin or its fragments play a role in the stimulation of IL-1 production, probably through monocytes adhering to the dialysis membrane. In addition to this dialysate factor, IL-1 production appears also to be stimulated by a blood-membrane interaction.