Fig. 4
A combination of Pam3CSK4 and PolyI:C (PP) recapitulates signatures of high nAb responses. a Heatmap showing cell activation, cytokines, and pathway signatures associated with nAb responses induced in human PBMCs by alum and TLR agonists. The data were selected from the experiment presented in Supplementary Fig. 5 and compared to the same changes triggered by YF-17D in the same donor PBMCs, presented in Fig. 1. b–g Immune stimulation by individual and combination of Pam3CSK4 and PolyI:C in human PBMCs. b, c MFI of activation markers on mDC-1 and monocytes. Antibodies against activation markers were labeled with the same fluorophore. d, e Levels of IL-12 and IFN-γ in the supernatant. f, g Induction of IFN-stimulated IFIT1 and OAS2 mRNA by quantitative real-time RT-PCR, normalized to GAPDH. h–j Activation of reporter genes in reporter cell lines. h Schematic reporter constructs in THP-1 and Jurkat cell lines. i, j Induction of NF-κB and ISRE reporter in THP1 and Jurkat cells. Data shown are the mean ± SEM for four independent PBMC donors or two independent experiments in triplicate for cell lines. Significant differences are determined using a two-tailed paired t test between treatment and mock. *p < 0.05, **p < 0.01, ***p < 0.005
