Figure 6. Co-expression of CS protein and EAT-2 increases cytolytic activity of CS protein specific T cells.

Co-vaccination of mice with Ad-CSP and Ad-EAT2 increased specific killing cells pulsed with CS protein peptides. BALB/cJ mice (n = 4) were co-injected IM with either 1×10⁸ vps/mouse Ad-CSP and 1×10⁸ vps/mouse Ad-GFP or 1×10⁸ vps/mouse Ad-CSP and 1×10⁸ vps/mouse Ad-EAT2 on Day 0. Day 14 splenocytes were collected from naïve mice and pulsed with either NYDNAGTNL peptide or an irrelevant peptide. NYDNAGTNL pulsed splenocytes were stained with a high concentration of CFSE while splenocytes pulsed with irrelevant peptide were stained with a low concentration of CFSE. Stained splenocytes were then combined in equivalent doses. 8 million cells were then injected IV into naïve, Ad-CSP+Ad-GFP co-vaccinated, or Ad-CSP+Ad-EAT2 co-vaccinated mice. After 18 hrs splenocytes from these mice were collected and analyzed by flow cytometry to assess the amount of NYDNAGTNL specific killing. % Specific killing  =  1-((%CFSE^High/%CFSE^Low)_immunized/(%CFSE^High/CFSE^Low)_{non-immunized}). * denotes significant difference between treatments p<0.05.