Figure 2.

Proliferation and cytokine production of CD4⁺ T cells after stimulation with ovalbumin (OVA) or advanced glycation endproduct (AGE)-OVA loaded mature dendritic cells (DCs). (a) Autologous T cells were cultured together with mature DCs pulsed with different concentrations of OVA or AGE-OVA or tetanus toxoid (TT) as a positive control. For the negative control (medium) no allergen was added. After 5 days, the cells were pulsed for 8 hr with [³H]thymidine. (b) DCs were pulsed with different concentrations of OVA or AGE-OVA or TT during their maturation phase from day 6–8. Then, interleukin (IL)-6 and IL-12p40 production was determined by enzyme-linked immunosorbent assay (ELISA). (c) CD4⁺ T cells were cultured together with allergen-loaded mature DCs. After 8 days the co-cultures were re-stimulated with freshly prepared autologous allergen-loaded DCs. The concentrations of cytokines were determined after 24 hr by ELISA. Shown is the mean ± standard deviation of five (for b) and 10 (for a and c) independent experiments; *P ≤ 0·05 compared with OVA. (d) Co-cultures were further stimulated with phorbol 12-myristate 13-acetate (PMA)/ionomycin/GolgiStop™ for 5 hr and stained for intracellular IL-4 and interferon (IFN)-γ. One representative of four independent experiments is shown.