Fig. 5

Keratinocyte-initiated Th1 differentiation is dependent on costimulation through CD58/CD2. Naive CD4⁺ T cells were cultured for the indicated time periods with untreated KCs (white bars) or IFNγ-pretreated KCs (black and gray bars) loaded with SEB (+) or not (−). Cytokine expression in the T cells was analyzed by flow cytometry, or cytokine secretion into the supernatants was determined by a cytokine bead array. Effects of isotype control antibodies (Iso), blocking antibodies against CD58, or CD2 downmodulation (CD2mod) in T cells on intracellular IFNγ expression (a), TGFβ expression (b), and IL-6 secretion into the supernatant (c) after 24 h of coculture (n ≥ 6 individual T cell donors). Effects of isotype control antibodies (Iso), blocking antibodies against CD58, or CD2 downmodulation (CD2mod) in T cells on the differentiation of Th1 (IFNγ⁺) cells (d), Th1 (CD183⁺CD194⁻) cells (e), and Th17 (IL-17A⁺) (f) after 6 days of coculture. Intracellular cytokine expression and CD183 and CD194 surface expression were analyzed after PMA/ionomycin treatment (n ≥ 6 individual T cell donors). Effects of exogenously added recombinant IFNγ (rIFNγ) on Th1 (IFNγ⁺) (g) and Th17 (IL-17A⁺) (h) differentiation after CD2 downmodulation (CD2mod) in a 6-day coculture (n = 4 individual T cell donors). Intracellular cytokine expression was analyzed after PMA/ionomycin treatment. Data are represented as the mean ± SEM. ***p < 0.001; **p < 0.01; *p < 0.05; and ns not significant. See also Fig. S5