In situ hybridization with 35S-labeled Epstein-Barr virus (EBV) probes was applied to paraffin sections of tonsils from seven patients with clinical, serologic, and morphologic evidence of acute infectious mononucleosis. EBV genomes were demonstrated in activated lymphoid B blasts in the interfollicular and perifollicular zones in all these cases. However, in no case could EBV be identified in epithelial cells. These results are at variance with the current concept which attributes a central role to the tonsillar epithelium in primary EBV infection.