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. 2020 Jan 9;11:147. doi: 10.1038/s41467-019-13837-4

Fig. 5. TLR8 enhances differentiation of CD4+ T cells towards Th1 and Th17.

Fig. 5

a Purity analysis of CD4+ T cells after isolation and content of contaminating CD14+ and CD11c+ cells. b, c CD4+ T cells were TCR-activated in the presence or absence of 5 μg/ml CL264 (TLR7), R848 (TLR7/8), CL75, pU/pLA (TLR8), or CpG (TLR9, 5 μM). Production of CD4+ T helper effector cytokines IFN-γ, IL-17, IL-4, TNF-α, and IL-2 was analyzed at 24–144 h by intracellular flow cytometry. b Example of flow cytometric analysis of Th1 lineage cytokine IFN-γ and Th17 cytokine IL-17 at 48 h. c Frequencies of cytokine-producing CD4+ T cells over time. Results represent mean + SEM from eight independent experiments. d CD4+ T cells were pre-treated with TLR8-specific inhibitors CU-CPT9a, CU-CPT9b, or the negative control compound CU-CPT6 for 2 h prior to TCR activation and stimulation with 5 μg/ml of CL264, CL75, pU/pLA, or FSL-1 (TLR2) for 72 h. Cytokine production was analyzed by intracellular flow cytometry. Results from inhibitor-treated CD4+ T cells were compared to untreated (−) or DMSO (=0) treated control cells (same data in all three graphs for each cytokine). Bars represent mean + SEM from ten experiments. Statistical significance was determined from log-transformed data by repeated measures two-way ANOVA with Dunnettʼs post-test in c (48 h timepoint) and in d; significance levels: *p < 0.05; **p < 0.01; ***p < 0.001. Source data are provided as a Source Data File.