HEp-2 cells, human epithelial cells derived from a larynx carcinoma, were found to be highly susceptible to infection with HIV-1 stain IIIb and MN, but not to infection with the monotropic strain IIIBa-L or the clinical isolate HIV-1AT. HEp-2 cells infected with HIV-1 IIIb continuously secreted high levels of p24 antigen, while no cytopathic effects were observed. Although no CD4 antigen could be detected on the cells by flow cytometric analysis, CD4 mRNA was detected by reverse transcriptase PCR. Furthermore, infection could be blocked by anti-CD4 monoclonal antibody OKT4a indicating a CD4 mediated viral entry in HEp-2 cells. HEp-2 cells are commonly used in clinical virology for the culture of different viruses from clinical specimens. HEp-2 cells should therefore be handled with caution as they may potentially become infected with HIV.