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. 2014 Feb 5;9(2):e88327. doi: 10.1371/journal.pone.0088327

Figure 3. Expression of constructs and T cell responses.

Figure 3

A) Human moDCs were transduced with either the non-ubiquitinated (0x) or ubiquitinated (1x) HIV-1 ZM96 gag constructs for 24 h or untransduced (control). Cells were then cultured for a further 18 h in the absence (hatched bar) or presence (grey bar) of the proteasome inhibitor, MG132. Cells were then harvested and stained intracellularly with anti-gag-PE antibody and analysed by flow cytometry. Results shown are mean±SD for 3 donors. B) MoDCs were transduced with either the non-ubiquitinated (0x, hatched bar) or ubiquitinated (1x, grey bar) HIV-1 ZM96 gag constructs and co-cultured with autologous naïve T cells. After 4 weeks of T cell expansion in culture, T cells were harvested and restimulated with single individual peptides and IFNγ production was determined by ELISPOT. Overall fusion with ubiquitin lowered the response although responses to 7 peptides were increased. For the 3 donors not shown only 2/49, 5/49 and 1/49 peptides gave a better response with Ub-gag. Graph represents 1 of 4 donors.