Figure 6

Vpx does not enhance HTLV-1 transmission to MDM or MDDC. (A) Jurkat cells were cotransfected with an antisense-intron-containing HTLV-1 or HIV-1 luciferase reporter plasmid, the respective packaging plasmid and pVSV-G. After 24 h, the transfected Jurkat cells were cocultured at a ratio of 2:1 with MDM that had been preincubated for 2 h with 25 ng p27 of Vpx-containing (+Vpx) or control VLP (−Vpx). After 48 h, the Jurkat cells were removed and adherent MDM were lysed and the luciferase activity was determined in quadruplicates. Background luciferase activity was determined by transfection of Jurkat cells with control plasmids (mock). The data are expressed as relative light units (RLU). One of four independent experiments is shown. (B) Jurkat cells were cotransfected with HTLV-1 or HIV-1 GFP reporter plasmids, the respective packaging plasmid and pVSV-G. After 24 h, the transfected Jurkat cells were cocultured at a ratio of 2:1 with MDDC that had been pretreated with 25 ng p27 of Vpx-containing (+Vpx) or control VLP (−Vpx). After 48 h, the cells were analyzed by flow cytometry to detect transmission of HIV-1 GFP and HTLV-1 GFP from the Jurkat cells to the MDDC by staining with anti-CD3-PECy5 to detect the Jurkat cells and anti-CD11c-APC to detect the MDDC. The infected MDDC were defined as the CD3-/CD11c + population. The data are the average of triplicate infections with error bars to indicate the standard deviation. One of three independent experiments is shown.