The human immunodeficiency virus type 1 (HIV-1) is rarely transmitted through salivary secretions, due in part to the presence of endogenous inhibitors. Here, the protective characteristics of the intraoral environment are summarized and inhibitory factors that reduce HIV-1 infectivity in vitro described, focusing on secretory leucocyte protease inhibitor (SLPI), a 12-kDa mucosal protein that blocks HIV infection in several cell-culture systems. SLPI appears to interact with a cellular surface molecule to limit viral entry into target cells. To determine whether the inhibitor has a similar role in vivo, the contribution of salivary SLPI to anti-HIV-1 activity was assessed. Whole unstimulated filtered salivas from infected and uninfected donors contained similar concentrations of the inhibitor. Depletion from SLPI filtered saliva produced a corresponding loss of inhibitory activity. In general, filtered whole salivas obtained from 10 donors had antiviral activities that correlated positively with SLPI concentrations. However, some samples having SLPI well below the concentration required for inhibitory activity in vitro exhibited modest inhibition, suggesting the presence of other anti-HIV-1 components in oral fluids. Thus, SLPI is a major but not sole inhibitor of this virus in saliva.