To elucidate further the influences of dietary fat on autoimmune diseases, two groups of NZB/W F1 mice were fed with diets containing 200 g dietary fat/kg and 50 g dietary fat/kg (control) respectively. The difference in energy intake between these two groups was compensated with carbohydrate. Mice were bled regularly every month and some of them were killed for in vitro experiments after 5 months experimental diets. Higher immunoglobulin (Ig)M and IgG anti-double stranded DNA antibody levels, shortened life span and worsened proteinuria were noted in mice fed on the high-fat diet compared with those fed on 50 g dietary fat/kg. Phenotypic analyses of spleen cells and peritoneal exudate cells showed that the percentage of CD5+ B cells and the mean fluorescent intensity of major histocompatibility molecules on the surface of both types of cells were higher in mice fed on the high-fat diet. In general, higher type 2 T-helper cell activity was noted in mice fed on the high-fat diet. In addition, cytokines such as interleukin-6, tumour necrosis factor-alpha and prostaglandin E2 (PGE2) produced by lipopolysaccharide-stimulated peritoneal exudate cells were also higher in the high-dietary-fat group. These studies suggest that high dietary fat and its related PGE2 level might have a critical effect on the frequency of CD5+ B cells, cytokine production, macrophage function and subsequent autoimmune regulation in autoimmune mice.