Fig. 5.

Anti-ILT4 enhances the generation of polyfunctional CD8⁺ T cells. (A) Dermal CD14⁺ DCs were cultured at a ratio of 1:40 with allogeneic CFSE-labeled naïve CD8⁺ T cells and 20 µg/mL indicated anti-ILT4 mAb (clones 20F3, 17E5, and 8E10) or an isotype-matched control. After 9 d, the cells were assessed for their proliferation based on the dilution of CFSE dye. Graph shows mean results of two independent experiments. (B) CD8⁺ T cells primed for 11 d by dermal CD14⁺ DCs in the presence of indicated anti-ILT4 mAb or an isotype-matched control were sorted as CFSE^loCD11c⁻CD4⁻ cells and cultured at 1.5 × 10⁵ cells per mL. IL-13 production was measured in the culture supernatant by using Luminex following 48-h stimulation with anti-CD3 and -CD28 mAbs. (C) CD8⁺ T cells primed for 9 d by dermal CD14⁺ DCs in the presence of anti-ILT4 (20F3) were expanded with anti-CD3 and -CD28 mAbs and IL-2 for 48 h and assessed following 5 h of reactivation with PMA and ionomycin for the intracellular expression of IFN-γ, TNF-α, and granzyme B by flow cytometry.