Figure 8

HIV-1 replicates in unstimulated CD4⁺T lymphocytes treated with exosomes from Nef _F12 -expressing cells. A. Detection of Nef in cells expressing either Nef_F12 or Nef_G2A, and in exosomes purified from the respective supernatants. Shown is the anti-Nef western blot analysis of both cells and exosomes from either 293 T cells transiently transfected with vectors expressing Nef_F12 or Nef_G2A, or mock-transfected cells (Ctrl). Signals from cellular Nef were normalized with β-actin detection, and anti-ICAM-1 analysis served to normalize exosome signals. On the left of Nef-specific panels, arrows indicate the specific signals. On the right, molecular markers are given in kDa. Results are representative of two independent experiments. B. HIV-1 replication in unstimulated CD4⁺ T lymphocytes treated with exosomes from cells expressing either Nef_F12 or Nef_G2A. 10⁵ unstimulated CD4⁺ T lymphocytes were challenged with 60 μU of exosomes from either F12/Hut-78 cells, 293 T cells transfected with Nef_F12- or Nef_G2A-expressing vectors, or mock-transfected cells (Ctrl), and then infected with 50 ng of HIV-1. As control, unstimulated cells were challenged with HIV-1 alone. Three days later, the cells were analyzed by FACS for HIV-1 expression. The fold increases of the percentages of HIV-1 CAp24-positive cells compared to cultures treated with HIV-1 alone are reported. Shown are the mean of fold increases + SD as calculated from three independent experiments with duplicates. *p < 0.05.