(A) PBMC were pre-treated with TLR8 antagonist or control for 30m before treatment with TLR2/1L, ssRNA and mTB tRNA. Cytokine secretion was measured at 24h. (IL-18 and IL-6: (n=3); IL-12p70 and IFN-γ (n=5). Statistical significance calculated by two-way repeated measures ANOVA and multiple comparisons by Tukey post-hoc test. * P < 0.05, ** P < 0.01. (B) PBMC were stimulated with nucleotide oligomers complexed with Dotap (endosomal PRR) or Lipofectamine2000 (cytosolic PRR) or synthetic agonists for 24h (n>=4). Statistical significance of ligand-stimulated PBMC vs. media calculated by one-way repeated measures ANOVA followed by Dunnett’s multiple comparisons test. (C) PBMC were stimulated with poly I:C, TL8-506 or combination and cytokine secretion measured at 24h (n=3). Data summarized using a mixed-effects model with a fixed effect for ligand and a random effect for subject. A two-way interaction term was fit to each ligand to test synergy. The likelihood ratio test was used to test for statistical significance. Data are represented as mean ± SEM. *p<= 0.05, ** p<=0.01, *** p<=0.001