Figure 4

NNRTI induced selective killing of HIV-1 infected primary human cells. (A) PBMC prepared from buffy coats of healthy blood donors were infected with HIV-1AGFP. At day 2 post infection, 100 ng/ml AMD-3100 was added to all samples to prevent further infection. Individual samples were incubated in addition with DMSO (white bars), 200 nm DRV (gray bars), 1 μM of the indicated NNRTI (black bars) or 1 μM NNRTI + 200 nM DRV (hatched bars), respectively. After further incubation for 5 days, cells were harvested and analyzed for the proportion of infected GFP expressing cells by flow cytometry. The figure shows mean values and standard deviations from three independent experiments (VRX-480773) or one experiment (GW-678248), respectively, each comprising three parallel cultures using different donor pools. P-values were calculated using a two-tailed unpaired t-test (GraphPad Prism). Values were normalized to the respective solvent control. (B) CD4 positive cells isolated from PBMC were infected with HXB2D-EGFP. At day 7 post infection 1 μM AZT (white bars), 1 μM of the indicated NNRTI (striped bars), 1 μM of the indicated NNRTI + 1 μM AZT (black bars) or 1 μM NNRTI + 1 μM AZT + 100 nM DRV (hatched bars), respectively, were added. After further incubation for 3 days, cells were harvested and analyzed for the proportion of infected cells by flow cytometry. The figure shows mean values and standard deviations of values from one representative experiment (three parallel infections). Asterisks: non-infected controls.