LMP1 induces enhanced T cell responses in a Gag peptide-specific IFN-γ ELISPOT assay. (A) Schematic of the experimental protocol. DCs from an HIV seronegative donor were transduced with scSIV viruses for 4 days, washed, and then incubated with autologous T cells for 12-days in the presence of nevirapine and IL-2 (5 U/ml) starting on day 3 of the coculture. After 12 days, cultures were restimulated with a consensus SIVmac239 15-mer Gag peptide pool and IFN-γ ELISPOT analysis was performed 24 hours later. (B) DCs infected with parent scSIV were unable to stimulate anti-SIV T cell responses, while the nef-deleted virus scSIV-GFP induced a modest T cell response. DC infected with scSIV-LMP1 and LMP1-CD40 significantly enhanced anti-Gag T cell responses (p < 0.001). Results are representative of three independent experiments using three different donor blood samples.