The association between sexual activity and human herpes virus-8 (HHV-8) infection has been established, but the mode of acquisition is still unclear. Blood samples from 238 individuals from Northern Cameroon were tested to evaluate the incidence of herpes simplex virus-2 (HSV-2), human immunodeficiency virus (HIV), Treponema pallidum, and HHV-8 infections and to identify their possible association. The presence of HSV-2 antibodies was associated significantly with gender, age, and HIV, HHV-8 antilatent, and T. pallidum antibodies, but not with HHV-8 antilytic antibodies. In a multivariate model older age, female gender, seropositivity for HIV, for HHV-8 latent antigens and for T. pallidum were associated independently with seropositivity for HSV-2. HSV-2-seropositive individuals had significantly higher titers of antibodies to both lytic (P = 0.019) and latent (P = 0.021) HHV-8 antigens. These results suggest that HSV-2 infection can contribute to sexual transmission of HHV-8 infection.