Figure 1. Ad-siA20-BM-DCs induce potent systemic HIV gp120–specific immune responses.

Groups of C57BL/6 mice were immunized with HIV gp120 protein–pulsed (100 μg/ml), Ad-transfected BM-DCs (1 × 10⁶ cells/mouse) twice with a week interval, followed by no stimulation or poly(I:C) (50 μg/mouse) or LPS stimulation (30 μg/mouse) (i.p.) daily for 3 consecutive days after each DC immunization. (A) CD8⁺ T cells or CD4⁺ T cells isolated from pooled splenocytes of immunized mice (n = 2–3) were subjected to IFN-γ ELISPOT assays. (B and C) ICS of CD8⁺ T cells (B) and CD4⁺ T cells (C) from draining LNs of immunized mice was performed after 6–8 hours in vitro restimulation. (D) The expression levels of perforin, granzymes A and B, and FasL in gated CD8⁺ T cells of pooled draining LNs of immunized mice are shown. (E) Systemic antibody responses enhanced by siA20-BM-DCs. HIV gp120–specific IgG1 and IgG2a subclass titers from the pooled sample of each group (4–6 mice/group) were quantified by capture ELISA. Antibody titers are reported as the mean ± SD of endpoint titers (36). Experiments were repeated 3 times with similar results. *P < 0.05, **P < 0.01, Ad-siGFP-BM-DCs versus Ad-siA20-BM-DCs.