Human herpesvirus 6 (HHV-6) is a beta-herpesvirus capable of infecting cells from different origin. In this study, infection with HHV-6A of human embryonic fibroblasts (HEFs) was performed. Infected cells showed obvious cytopathic effects (CPE). PCR and immunohistochemical tests also confirmed that HEFs are susceptible to HHV-6A infection. The biological effects of HHV-6A infection on HEFs were studied. Infected cells showed decreased proliferation as measured by [3H] thymidine incorporation and cell counting. Further analysis demonstrated that infection with HHV-6A leads to cell cycle arrest at G2/M phase and increasing cell death. This is the first demonstration that infection of HEFs with HHV-6A causes profound alterations of cell properties. J. Med. Virol. 84:657–663, 2012. © 2011 Wiley Periodicals, Inc.