Figure 5

HIV-2 ROD-GFP and MDDCs. (a) Susceptibility of MDDCs to HIV-2 ROD-GFP infection. MDDCs were exposed to HIV-2 ROD-GFP, pseudotyped or not with VSV-G (50 and 150 ng p27 mL-1, respectively). After 3 days, the levels of GFP were measured by flow cytometry. Results from 6 independent donors are shown (b) HIV-2 ROD-GFP binding. MDDCs were exposed to the indicated doses of HIV-2 ROD-GFP, pseudotyped or not with VSV-G. After 2 h at 4°C, cells were extensively washed and the amount of cell-associated p27 was assessed by ELISA. Data are Mean ± SEM of 4 independent donors. ns: non significant (c) HIV-2 ROD-GFP fusion. MDDCs were exposed to the indicated doses of HIV-2 ROD-GFP, pseudotyped or not with VSV-G, and bearing the chimeric protein β-lactamase-Vpr. After 2 h at 37°C and 2 h at room temperature, viral access to the cytoplasm was assessed by flow cytometry, using the ability of β -lactamase to cleave the cytoplasmic CCF2-AM fluorogenic substrate. A mean ± SEM of 4 independent donors is shown. *: p-value < 0.05. Comparisons were made between the condition indicated and the no VSV condition at the same viral inoculum. (d) HIV-2 ROD-GFP DNA synthesis. MDDCs were exposed to HIV-2 ROD-GFP pseudotyped or not with VSV-G, in the presence or absence of AZT. After 3 days, the cells were harvested for HIV-2 DNA quantification by qPCR. Data are mean ± SEM of 4 independent donors.