Fig. 3.

T helper responses indicated by cytokine Release of INF-γ, IL-5, IL-10 from splenocytes harvested a 7 days post prime-boost and b 21 days post prime-boost, restimulated in vitro with DCh5T4. Cytokine release determined from culture supernatant from splenocytes restimulated in vitro for 96 h with DCh5T4 at an E;T ratio of 20:1 was tested using the cytokine Elisa kits from BD Pharmingen. Early cytokine responses represent a mixture of Th1 and Th2 cytokines. Splenocytes vaccinated Adh5T4/Adh5T4 only secrete INF-γ, whilst splenocytes vaccinated with DC5T4/Adh5T4 or Adh5T4/DCh5T4 secrete all three cytokines. It is difficult to interpret cytokine profiles from the DCh5T4/DCh5T4 as there is non-specific release of cytokines with DC2.4 restimulation in vitro. 21 days following boost there is no IL-10 secretion by splenocytes. IFN-γ secretion is no longer detectable in the Adh5T4/Adh5T4 vaccination group. Interestingly the mixed Th1/Th2 cytokine profile is sustained with DCh5T4/Adh5T4 vaccination, whilst Adh5T4/DCh5T4 appears polarised to a Th1 biased response. (1 Adh5T4/Adh5T4 vs. AdGFP/AdGFP, 2 DCh5T4/DCh5T4 vs. DC2.4/DC2.4, 3 Adh5T4/DCh5T4 vs. AdGFP/DC2.4, 4 DCh5T4/Adh5T4 vs. DC2.4/AdGFP)