Figure 4.
Immune-complexed IgG-OVA facilitates DC-mediated activation of OVA-specific OT-1 CD8 T cells. Ovalbumin (0.1, 1, 10, and 100 μg/mL) was preincubated for 10 min either alone or in the presence of ovalbumin-specific Abs (1, 10, and 100 μg/mL) to generate OVA-ICs. OVA or OVA-ICs were then loaded to 5 × 105 bmDCs and incubated overnight at 37 °C. Then, bmDCs were washed and cocultured with 5 × 105 isolated OVA-specific CD8 TCR tg OT-1 T cells for 48 h. After 48 h of co-cultivation, activation of CD8 T cells was determined by the expression of CD25 and the early activation marker CD69 gating on AAD-negative living CD8 singlets measured by FACS. Data represent mean ± SEM of three independent experiments. Data were analyzed by GraphPad PRISM software using ANOVA followed by Dunnet’s multiple comparison test (***, **, * significant at p < 0.001, p < 0.01, p < 0.05, respectively).