Figure 4. Cross-reactive cell-mediated immunity in epitope modified minigene immunized HHD mice. HHD mice (n = 5 per group) were immunized by DNA electro-gene transfer with candidate epitope modified minigenes (EMMs; refer to Figure 3) receiving 4 weekly subcutaneous injections of 50 μg plasmid DNA followed by electroporation. Two weeks after the 4th injection, peripheral blood mononuclear cells (PBMCs) from mice within the indicated group were combined and challenged in vitro using the pool of analogs or wild-type (wt) epitopes. Cell-mediated immunity was measured by CD8⁺ T cell IFNγ secretion by intracellular staining of cell-surface stained PBMCs and flow cytometry. The experiment was performed in duplicate and average values are reported.