Figure 2

HIV-1 access to DC cytosol is a prerequisite for HIV-1 impairment of the ability of DCs to prime T_N-cell responses. HIV-1 _BaL or HIV-1gp120_BaL protein was pretreated with anti-gp120 mAb (b12) for ∼60 min at 37°C. The matured DCs were treated with mock, mock-b12, HIV-1, HIV-1-b12, HIV-1gp120, HIV-1gp120-b12, HIV-1-HIV-1gp120, and HIV-1-b12-HIV-1gp120-b12. DCs were also pulsed with fusion inhibitor (C34), or in combination with b12 antibody. The DCs were pulsed for ∼ 24 h, washed and cocultured with CFSE-labeled T_N cells. DC-T-cell cocultures were restimulated on d 7 and harvested on d 8. (A, B) T-cell proliferation was measured for CFSE dilution by flow cytometry and ³H-thymidine incorporation by β counter. (C) The gene profiles of CTLA-4, PD-1, TRAIL, TIM-3, CD160, LAG3, BLIMP-1, FOXP3 and DTX1 were analyzed by qRT-PCR (n = 5) in an assay with or without b12 mAb. The experiments (A, n = 5, and B, C, n = 8) were normalized and statistical analysis was performed by one-way ANOVA (Tukey post hoc test).**P < 0.005, ***P < 0.001. (D) Protein expression profiles of CTLA-4, PD-1, TRAIL, TIM-3, CD160 and LAG3 were measured by flow cytometry and shown by one representative of 4 experiments (n = 4).