Fig 3. Bystander DC cytokine transcription profile.

A) Transcription profile of selected cytokines from bystander DCs at 12 h post-activation following exposure to HAdV5- or IC-HAdV5-challenged direct DCs. The transcription profile of bystander DCs created by IgG-challenged direct DCs was used as the baseline. For the genes in bold, primer sequences were designed in-house (n = 2 donors). B) Principal component analysis (PCA) of the changes in the 66 mRNAs included in the array. Three principal components showed 61% (F1), 28% (F2), and 11% (F3) accordance. C) TNF, IFNβ, and CXCL10 mRNA levels in bystander DCs. Direct DCs were mock-, IgG-, LPS-, HAdV5-, or IC-HAdV5-challenged. IC-HAdV5s were used at 20 x 10³, 10 x 10³, 5 x 10³, or 1 x 10³ physical particles/direct DC. Assays were carried out in 3 donors in at least duplicates. The fold increase is shown as mean ± SEM. See S1 Table for additional statistical analyses. D) Kinetics (3, 6, 18 h) of TNF (top panels) and IFNβ (bottom panels) mRNA levels of THP-1-derived DC challenged with LPS, HAd5, IgG or IC-HAdV5 (left panels) or bystander DCs (using the milieu from the direct DCs in the left panel) (right panels). Three independent assays in duplicate were performed. The fold increase is shown as the mean ± SEM. P values were derived from one-way ANOVA with Dunnett’s test: ** p < 0.01 and *** p < 0.001.