The prevalence and incidence of human immunodeficiency virus types 1 and 2 (HIV-1, HIV-2), human T-lymphotropic virus types I and II (HTLV-I/II), and syphilitic infections and the association between these infections were determined in a cohort of police officers in Guinea-Bissau. Between January 1990 and December 1992, 1,384 subjects (1,241 men and 143 women) were included in the study; and of the first 879 tested, 561 were tested at least for a second time. The overall seroprevalence of HIV-1 was 0.4%, of HIV-2 11.6%, and of HTLV I/II 4.4%. Three individuals (0.2%) were seropositive for both HIV-1 and HIV-2. Women had a significantly higher prevalence of HIV-2 infection than men (16.8% and 11.0%, respectively, p < 0.05). Serologic evidence of previous syphilis was present in 12.1% and was significantly more common in men (12.7%) than in women (7.0%) (p < 0.05). There was a significant association between the prevalence of HIV-2 and HTLV-I/II infection (p < 0.05). The annual incidence of HIV-1 was 0.7%, of HIV-2 1.6%, of HTLV I/II 0.4%, and of syphilis 1.7%. There was no association between the incidence of the various infections. The death rate (per 100 person-years) was significantly higher among HIV-2-infected individuals (2.7%) than among HIV-negative individuals (0.5%) (relative risk = 5.1; 95% confidence interval, 2.1–12.2; p < 0.001). HIV-related symptoms were more frequent among the HIV-2-positive individuals who died compared with the seronegative individuals who died (p < 0.01). In conclusion, the prevalence and incidence of HIV-2 infection were high, whereas the incidence of HIV-1 infection was disproportionately high in relation to the low prevalence, indicating a recent increased spread of HIV-1 infection in Guinea-Bissau.