FIG 5.

Group O Nefs cannot attenuate innate sensing of infected cells by pDC-containing PBMCs. (A) Schematic representation of the experimental system. WT or dU HIV-infected T cells were cocultured with freshly isolated PBMCs, and levels of bioactive IFN-I released into the supernatants were measured 18 to 24 h later. (B and C) MT4 T cells were mock infected or infected with GFP-expressing NL4.3 WT or dU viruses expressing either NL4.3 Nef, M-MRCA Nef, O-Nefs, or N-MRCA Nef, as indicated, and cell populations harboring similar percentages of infected T cells were cocultured with PBMCs. After 24 h of coculture, levels of IFN-I released into the supernatants were measured. Representative examples of absolute levels (B) and relative percentages (C) of IFN-I detected after coculture of HIV-1-infected MT4 donor cells with PBMCs (n = 8) are shown. The amount of IFN-I released by PBMCs in contact with dU/NL4.3 Nef-infected cells was set at 100%. Repeated-measures ANOVA with Bonferroni's multiple-comparison test was used (***, P < 0.001; ns, not significant [P > 0.05]). Error bars represent standard deviations.