HIV-1 Nef Impairs Key Functional Activities in Human Macrophages through CD36 Downregulation
Figure 9
Recombinant Nef reduces phagocytosis in MDMs.
PBMCs were cultivated in HEMA condition w/o EPO for three days followed by additional three days in presence of 50 ng/mL rNef/myr. Cells were then incubated with FITC-labeled beads or GFP-S. typhimurium for 30 min. (A) Representative dot plots of fluorescent beads and S. typhimurium uptake by Nef-treated (+Nef) compared to untreated (Ctr) cells evaluated by flow cytometry. Cells not incubated with beads or S. typhimurium were used as control for auto-fluorescent signal. The gates indicate the respective percent of phagocytosis. (B) The phagocytosis capability of Nef-treated (Nef) expressed as percent of control (Crt) is reported in the histogram. Where required by experimental procedures, control cells were pre-incubated with blocking anti-CD36 antibody for 20 min before the phagocytosis assay (Crt+anti-CD36). SYTOX Blue was used to exclude dead cells. The results (mean ± standard deviation) are representative of four independent experiments (*p<0.05).