MoDCs were transduced with either the non-ubiquitinated (0x, hatched bar) or ubiquitinated (1x, grey bar) HIV-1 CN54 gag constructs and co-cultured for 7 days with autologous T cells for cells from A) HIV infected patients or C) for healthy control donors. T cells were expanded in culture for 4 weeks, harvested and restimulated with peptide pools and IFNγ production determined by ELISPOT. In addition, mice were injected with either construct and one week post injection, spleens were harvested, stimulated with peptide pools and IFNγ production was determined by ELISPOT (B). IFNγ production by T cells from healthy human controls was also determined by intracellular cytokine staining of CD8 or CD4 T cells (D and E). Graph A) represents one donor of 3± SD, B) represents 3 mice±SD and is representative of one experiment out of three, C) represents 2 individual donors out of 6, D) represents 4 donors±SD and E) is a representative CD8+ intracellular flow cytometry plot.