Among the various mechanisms proposed to explain the pathogenesis of cerebral lesions in human immunodeficiency virus (HIV)-induced encephalitis, a cytokine-mediated action has found most favour. Indeed, elevated expression of cytokines such as interleukin (IL)-1 and tumor necrosis factor-alpha (TNF-alpha), thought to be neurotoxic, has been found in AIDS patients. As a previous study had demonstrated the presence of HIV proviral DNA in brain tissue of a number of HIV-positive non-AIDS patients, we undertook this present investigation using morphological, immunohistochemistry (IHC) and polymerase chain reaction (PCR) methods to detect the expression of major histocompatibility complex (MHC) class II molecules, the presence of HIV-1 proviral DNA and of the cytokines TNF-alpha, IL-1 alpha, IL-4 and IL-6 in brains of the same group of individuals. The study included brains of 36 asymptomatic HIV-1 positive patients and the results were compared with those of AIDS patients either affected by HIV encephalitis (n = 8) or exempt from any neuropathological changes (n = 10) as well as of normal controls (n = 5). Results show that: HIV proviral DNA could be detected by PCR in 17 out of the 36 brains from HIV-positive pre-AIDS cases; most (15 of 17) of PCR-positive brains showed minimal to severe expression of MHC class II antigen; and cytokines could be detected predominantly within white matter even at this early stage. The data demonstrated that the state of immune activation described in AIDS is already present at the pre-AIDS stage and suggest that the presence of cytokines may already trigger the cascade of events leading to brain damage.