Figure 8
Assessment of alveolar macrophage (AM) function based on the HIV infection status of individual cells. Adherent AMs were incubated with reporter beads to measure phagocytosis, superoxide burst, and bulk proteolysis. The cells were then stained with HIV-gag Quasar 670-labeled probes by fluorescence in situ hybridization (FISH) to detect HIV-infected AMs and analyzed by flow cytometry. (a) Representative flow cytometry pseudo-color plots showing phagocytosis of beads by small and large, HIV-infected, and HIV-uninfected AMs. The plots show four different cell populations: HIV-infected cells without beads (Q1), HIV-infected cells with beads (Q2), HIV-uninfected cells with beads (Q3), and HIV-uninfected cells without beads (Q4). (b) Phagocytosis of reporter beads by HIV-infected and HIV-uninfected AMs from the same HIV-infected individuals (n=14). (c,d) Phagosomal superoxide burst and bulk proteolytic activities, respectively, in HIV-infected compared with HIV-uninfected AMs (n=10). The data in panels c and d are expressed as the Activity Index, which is the ratio of relative fluorescence units of the substrate fluorescence divided by the calibration fluorescence to ensure dosage correction for minor variations in bead number. Data were analyzed using matched-pair one-way analysis of variance with Bonferroni correction; black horizontal bars represent medians (b) and means (c,d).
