Figure 4

BST-2/tetherin impact on DC-mediated HIV-1 transfer in trans. (A). 1 × 10⁵ DC, previously transfected (or not) with siTetherin or si Control (siCtr) for 48 hours, were left untreated (NT) or treated with IFN-α or LPS and challenged for six hours with 200 ng P24gag of HIV-ΔVpu or HIVF522Y viruses. DC, extensively washed, were then co-cultured with 1 × 10⁵ autologous CD4⁺ T cells pre-treated 30 minutes before co-culture with Indinavir. HIV-1 transfer on autologous CD4⁺ T cells was scored by FACS analysis of CD3⁺p24gag⁺FITC cells, four days post-transfer. Data shown are representative of three independent experiments. (B). Fold enhancement of HIVWT and HIV-ΔVpu transfer in CD3⁺/CD4⁺ T cells. A value of 1 was assigned to the percentage of transfer in the non-treated (HIV) conditions. Error bars indicate standard error mean (SEM). Data are representative of four different experiments. P-values of paired bilateral Student Test were not significant (p>0.18). (C). DC were transfected with siTetherin or siControl (siCtr) and left untreated (NT) or treated with IFN-α for 24 hours. Tetherin expression was analyzed by western blot. Data are representative of four experiments.