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. 2016 Jun 17;100(5):855–864. doi: 10.1189/jlb.2HI1115-525R

Figure 4.

Figure 4

ADAM23 expression in DCs impairs production of TH cytokine production under polarizing conditions. ADAM23 knockdown or control DCs pulsed with OVA323–339 were used to stimulate naïve CD4+ T cell responders for 10–12 d under TH0 (IL‐2 cytokine), TH1 (IL‐2, IL‐12p70, and IFN‐γ cytokines with anti‐IL‐4 neutralizing antibodies), TH2 (IL‐2 and IL‐4 cytokines with anti‐IFN‐γ and anti‐IL‐12p70 neutralizing antibodies), or TH17 (IL‐2, TGF‐β, and IL‐6 cytokines with anti‐IFN‐γ, ‐IL‐4, and ‐IL‐12p70 neutralizing antibodies) conditions. Restimulation using respective OVA323–339‐bearing DCs was on d 5, supplemented with respective cytokine/neutralizing antibody cocktails. On d 10–12, cells were stimulated with PMA and ionomycin mitogens in the presence of Brefeldin A for 4 h before staining for IFN‐γ, IL‐4, IL‐17, CD25, Foxp3, and CD4. Cells were then analyzed by flow cytometry. Dot plots show (A) IL‐4 versus IFN‐γ‐, (B) IFN‐γ versus IL‐17‐, and (C) CD25 versus Foxp3‐gated CD4+ T cell subsets. Isotype controls were used to establish gating strategies for all flow cytometric analyses. Data are representative of 3 independent experiments.