Signatures of inflammatory gene transcripts in PBMCs after TLR adjuvant injection. RNA was isolated from PBMCs cryopreserved with TRIzol reagent at 3 hours and 1, 3, 6, and 14 days after TLR-L injection and analyzed by low-density array quantitative real-time PCR for a panel of 93 genes involved in immune responses. All gene analytes at all time points were first normalized to the average cycling threshold value of expression of the housekeeping genes for 18s ribosomal RNA, Actb (β-actin), and Gusb (β-glucuronidase). Transcripts were grouped by their immune function and origin and their expression was evaluated at the time points matching innate cells kinetics in blood. (A) Heat map of gene transcripts. Each row represents the fold change relative to baseline (day −12) at the indicated time point after TLR-L injection in each of the experimental groups. The 34 genes with at least a 2-fold change from baseline at any given time point in any of the experimental groups are presented. Kinetics of expression of ISGs (B): Isg-15, Ifit-1, Mx-1, and Oas-1; chemokines (C): Cxcl-10 (IP-10), Cxcl-11 (I-TAC), Ccl-3 (MIP-1α), Ccl-2 (MCP-1), and the B-cell growth factor Tnfrsf-17 (D) are represented as the relative change in mRNA copies at the indicated time points after TLR-L injection compared with baseline (day −12). Dotted line marks the cutoff (y = 1) of relative expression change. Data are means ± SEM. *P < .05; **P < .01; ***P < .001 by ANOVA.