The spinal cords from 15 patients with AIDS-associated vacuolar myelopathy (VM), 4 AIDS patients without VM, and 5 HIV-seronegative controls, were studied with immunocytochemistry for TNF alpha. CSF and blood from HIV-seropositive patients with VM (n = 16), non-vacuolar myelopathies (n = 8), CNS infection but no clinical myelopathy (n = 31), no clinical or radiological evidence of CNS disease (n = 9), and from 7 HIV-seronegative controls with motor neurone disease were assayed for TNF alpha using an ELISA technique. TNF alpha was present on immunostaining in all the 15 cords with VM studied. The stained cells were macrophages, microglia and endothelial cells. The amount of immunostaining was higher in cords with VM compared with cords from HIV-seropositive patients without VM (p = 0.001). The distribution of staining corresponded to the areas of pathology but did not correlate with the severity of the VM. Immunostaining was also higher in the HIV-seropositive group compared to the HIV-seronegative controls (p = 0.001). There was no significant difference in the levels of TNF alpha in the CSF of patients with VM compared to any of the other groups studied. Blood levels of TNF alpha were lower in the HIV-seropositive controls without CNS disease and in the HIV-seronegative MND controls, than in patients with VM, non-vacuolar myelopathies, and CNS disease. CSF TNF alpha levels did not appear to be a reliable indicator of intramedullary levels. The findings support the hypothesis that TNF alpha may be relevant in the pathogenesis of vacuolar change in VM.