Centripetal movement of phagosomes is inhibited in HIV-1–infected macrophages. (A and B) Primary human macrophages were noninfected (black bars) or infected with HIV-1_ADAWT (red bars) for 8 d. The cells were incubated for different time points with IgG-SRBCs at 37°C and then fixed and permeabilized. They were labeled and analyzed as described in Fig. 2. Peripheral SRBCs, situated at a distance to the nucleus of more than two SRBCs in diameter, were counted for at least 200 phagosomes per condition at each time point. Results are expressed as the percentage of total number of SRBCs; two independent experiments on different donors are shown. (C) Primary human macrophages were noninfected (black bars) or infected with HIV-1_YU-2WT (red bars) or HIV-1_YU-2ΔVpr (blue bars) for 8 d. Cells were incubated with IgG-SRBCs for 1 h at 37°C and then analyzed as in A. The means ± SEM of three independent experiments are plotted. (D) Primary human macrophages were infected with HIV-1Gag-iGFP for 8 d, then incubated with IgG-SRBCs at 37°C under a spinning disk confocal microscope equipped with 5% CO₂ and a heated chamber. Images were recorded every minute for 120 min. The distances covered by internal phagosomes were measured for 53 phagosomes in noninfected cells and 60 phagosomes in HIV-infected macrophages and were plotted against time. The speed was calculated by linear regression. (E) Primary human macrophages were treated as in C. Gallery of phase contrast images of noninfected (top panels; see also Video 1) and HIV-1Gag-iGFP–infected (bottom panels; see also Video 2) cells showing phagosome movement. Bar, 10 µm. *, P < 0.05; **, P < 0.005.