It has been reported previously that cells expressing a truncated form of CD4 which lacks the cytoplasmic tail of the molecule (truncation at position 402) were not sensitive to human immunodeficiency virus type 1 (HIV-1)-induced apoptosis in an acute-phase model of infection (J. Corbeil, M. Tremblay, and D. D. Richman, J. Exp. Med. 183:39–48, 1996). The role played by the cytoplasmic domain of CD4 in HIV-1-induced apoptosis was reexamined here with clones of A2.01 cells expressing different forms of CD4 and the DNA intercalant YOPRO-1 assay. Six days after virus exposure, we found evidence of apoptosis in A2.01 cells expressing the wild-type CD4 (A2.01/CD4), whereas enhanced apoptosis remained absent in cultures of A2.01/CD4.401 and A2.01/CD4.403 cells (A2.01 cells which express CD4.401 and CD4.403 molecules with truncations at positions 401 and 403, respectively). However, cell death by apoptosis measured with YOPRO-1 was found in cultures of A2.01/CD4.401 and A2.01/CD4.403 cells 15 days after virus exposure. This result was confirmed with a terminal dUTP nick end-labeling assay and propidium iodide staining. The long lag time postinfection required for apoptosis to be observed in cultures of infected cells expressing truncated forms of CD4 was due to the delayed viral replication in these cells, as shown by monitoring of the viral reverse transcriptase activity and HIV-1 p24 ^gag antigen expression. These results emphasize the relationship between virus replication and cell death by apoptosis.