Efficient HIV-1 transmission occurs following 1 hr exposure of the inner, but not outer, foreskin to cells highly infected with HIV-1. Such HIV-1-infected cells form viral synapses with mucosal/apical foreskin keratinocytes, leading to polarized budding of the virus. In inner foreskin, HIV-1 is in turn rapidly internalized by LCs that migrate to the epidermal-dermal interface. There, LCs form conjugates with T-cells thereby transferring HIV-1. Cell-free HIV-1 is inefficient at foreskin penetration. To mimic the in vivo mixture of genital fluids during woman-to-man HIV-1 sexual transmission, the effect of a mixture of seminal plasma from HIV-negative men mixed with cervico-vaginal secretions from HIV-positive women was investigated. Such mixture significantly reduced HIV-1 translocation in inner foreskin reconstructions.