Co-expression of CS protein and EAT-2 increases the breadth of response against CS protein.

Increased breadth of response against CS protein epitopes was observed in mice co-vaccinated with Ad-CSP and Ad-EAT2 as compared to the control vaccine. Splenocytes from groups of five BALB/cJ mice were collected and pooled together from 14 days post injection with either innate modulating treatments or control. ELISpots were performed to measure IFNγ secreting cells when stimulated with a CS protein peptide library made up of 52 15mers that overlap by 5 a.a. on either side. Wells that contained more than 15 spots were counted and compared between treatment groups (inset). (A) Mice were co-injected IM with 5×10⁷ vps/mouse of Ad-CSP and 5×10⁷ vps/mouse of Ad-GFP/rEA or 5×10⁷ vps/mouse of Ad-CSP and 5×10⁷ vps/mouse of Ad-GFP. (B) Mice were co-injected IM with 5×10⁷ vps/mouse of Ad-CSP and 5×10⁷ vps/mouse of Ad-EAT2 or 5×10⁷ vps/mouse of Ad-CSP and 5×10⁷ vps/mouse of Ad-GFP. As a negative conrol, naïve splenocytes were also tested against paired peptides from the peptide pool, with an average background of spots per paired peptides being only 2.2 spots.