Figure 4.

SMG DC are more effective in the activation of antigen-specific T cells than Static DC. JAWS II DC or BMDC (2 × 10⁵/ml) were cultured in Static (white and gray bars) or SMG (black bar) conditions for 72 and 48 hours, respectively. Some JAWS II DC (+cytokines) were incubated with a cocktail of cytokines (IFN-γ, IL-4 and TNFα) 6 hours prior to harvest. Subsequently, Static and SMG JAWS II DC were washed and incubated (1 × 10⁴/well) with OT-II CD4⁺ TCH (5 × 10⁴/well) and (a) OVA323-339 or (b) OVA protein for an additional 24 hours in Static conditions. Culture supernatants were collected and assessed for IL-2 production by ELISA. Wells containing Static or SMG JAWS II DC and OT-II TCH without OVA323-339 or OVA protein were not included in the figure and yielded 0 cytokine upon assessment. (c) OT-I CD8⁺ T cells (5 × 10⁴/well) were added to Static and SMG BMDC (1 × 10⁴/well) with or without OVA257-264 for 24 hours in Static conditions. Culture supernatants were collected and the production of IFN-γ determined by ELISA. For all panels, the data represents the mean + SD of quadruplicates of two independent experiments. In (a,b), *p-value ≤ 0.05 comparing the activation of T cell IL-2 production by SMG and Static JAWS II DC as well as SMG and Static JAWS II DC (+cytokines). In (c), *p-value ≤ 0.05 comparing the activation of T cell IFN-γ production by SMG and Static BMDC, both including OVA257-264.