Langerhans cells (LC) are bone marrow-derived, HLA-DR+, CD1a+, dendritic antigen-presenting cells found in stratified squamous epithelia. Within resident epidermal cells (EC), LC are the only cells expressing the CD4 antigen and are, therefore, a possible target for human immunodeficiency virus (HIV) infection. To date, conflicting results have been reported on the in vivo infection of LC by HIV. The aim of the present study was to investigate the presence of HIV-1 proviral DNA in epidermal LC of HIV-1-infected patients. EC suspensions were prepared from clinically normal skin of nine seropositive patients. Purified LC and LC-depleted EC were obtained by immunomagnetic separation and analyzed for the presence of HIV-1 proviral DNA by the polymerase chain reaction using primer pairs from different conserved regions (env and gag) of the HIV-1 genome. HIV-1 proviral DNA was detected in LC from seven of nine patients. LC-depleted EC fractions from the same nine patients were all negative, with the exception of one case. Altogether these results demonstrate that epidermal LC are infected by HIV-1 and constitute the only resident cell type in the epidermis harboring the virus. Further studies are, however, needed to demonstrate HIV replication in LC and to elucidate the functional role of LC in this infection.