FIG. 5.
Viral DNA in cells exposed to native and AT-2-inactivated HIV-1. Human 3-day PHA-induced lymphoblasts were exposed to HIV-1, and cell pellets were collected at 24 and 48 h. Total DNA from the cell pellets was prepared and tested by real-time PCR for reverse-transcribed HIV-1 gag and strong-stop DNA. Results were normalized based on copies of DNA coding for PBGD, as described in the text. Note the time-dependent accumulation of gag and strong-stop DNA in untreated controls and the absence of gag DNA in AZT- and ddI-treated cultures. AT-2 treatment of HIV-1 prevents the production of both gag and strong-stop DNA. Results shown are averages of duplicate determinations of the DNA copy number for each condition in one of three experiments with consistent results. In a separate experiment in which low levels of gag DNA were produced from virions treated with a suboptimal concentration of AT-2, no productive infection was observed (data not shown), suggesting that AT-2-treated virions may also be deficient in the ability to complete other post-reverse transcription, pre-integration steps of the viral life cycle in which p7NC has been implicated (26a).