Fig. 3. IL-6 secreted by cancer cells affects moDC-induced Th1 differentiation.

a MoDCs were differentiated and stimulated (LPS, 100 ng/ml, 48 h) in the presence or absence of 20% of NCI-H226 supernatant (SN). Stimulation of moDCs was additionally carried out in the presence or absence of tocilizumab (5 µg/ml, anti-IL-6R mAb). IL-12p70 secretion was analyzed by CBA assay. Shown are 2 independent experiments. b Same as in a, but moDCs were differentiated and stimulated (LPS, 100 ng/ml, 48 h) in presence of IL-6 depleted- or control- supernatant (SN) of NCI-H226. The relative IL12-p70 secretion is presented (mean fold change ± SD; n = 6). c A MLR was performed with allogeneic CD4⁺ T cells and moDCs differentiated and stimulated (LPS, 100 ng/ml, 48 h) in the presence or absence of IL-6 (20 ng/ml) or of 20% of NCI-H226 supernatant (SN). T cell proliferation was determined via ³H-thymidine incorporation. Shown is a representative experiment. Error bars show the standard deviation of three technical replicates. d The induced T cell proliferation of IL-6-treated moDCs (20 ng/ml) (mean fold change ± SD; n = 4) and of e, moDCs treated with 20% of NCI-H226 supernatant (SN) was determined relative to the corresponding stimulated control (mean fold change ± SD; n = 4). f Naïve CD4⁺ T cells were expanded by stimulation with allogeneic moDCs, that have been differentiated and stimulated (LPS, 100 ng/ml, 48 h) in the presence or absence of IL-6 (20 ng/ml) or of 20% of NCI-H226 supernatant (SN). After restimulation of the CD4⁺ T cells with accordingly treated moDCs CD4⁺ T cells were stained for their T-bet expression. Shown is the percentage of the T-bet⁺ population (mean ± SD; n = 3) and the relative mean fluorescence of T-bet (mean fold change ± SD; n = 3).