We investigated IFN-gamma, IL-4, IL-10 and TNF-alpha in the sera of 42 patients with demyelinating diseases: multiple sclerosis (MS) (21), Guillain-Barré syndrome (GBS) (14), Miller-Fisher syndrome (MFS) (7) during the acute or active stage and 12 normal controls using an originally devised sensitive sandwich enzyme-linked immunosorbent assay (ELISA). We found elevated serum levels of IFN-gamma derived from T helper 1 (Th1) cells and of IL-4 derived from Th2 cells in MS, GBS and MFS. It has been reported in several studies that IL-4 production is increased in MS; however, there have been no reports of elevated serum IL-4 levels in MS. We here describe elevated serum levels of IL-4 in MS, GBS and MFS for the first time. Serum TNF-alpha levels were significantly elevated in patients with MS (p < 0.01). Serum TNF-alpha levels also tended to be elevated in patients with GBS and MFS, but not significantly elevated compared with those of controls. Serum IL-10 levels were not elevated significantly in these diseases, although several patients had higher levels of serum IL-10 than controls. These findings suggest that simultaneous activation of both Th1 and Th2 cells may occur during the acute stage, indicating systemic involvement of the immune system in patients with MS, GBS and MFS. Levels of serum IFN-gamma were two- to three-fold higher than those of serum IL-4 in all three demyelinating diseases. We may then conclude that Th1 response may be dominant in GBS and MFS during the acute stage.