Extended Data Fig. 3: ALVAC augments CREB1 driven gene expression by 24 hours post immunization and CREB1 signaling increases up to 72 hours post vaccination.

a, To more clearly understand the temporal nature of ALVAC induced CREB1, we accessed data from samples taken from the P162 study in which gene expression was quantified at 16, 24, 48 and 72 hrs after each of the 4 vaccination timepoints (n = 6 animals). GSEA using the CREB1 gene set from ChEA was performed to identify the temporal kinetics of ALVAC induced CREB1 by quantifying the fold induction of genes compared to pre-vaccination. In general, CREB1 target genes were suppressed at 16 hrs and induced at 24, 48 and 72 hrs. b, To determine if the same genes which were downregulated in some timepoints overlapped with those induced at others, a network was generated using Jaccard index to determine overlap in the CREB1 gene set leading edges for each of the timepoints. Red node color indicates induced, blue node color indicates suppressed. Node edge is colored per timepoint. Thickness of lines connecting nodes reflects the Jaccard index such that the thicker the line the more overlap. As can be seen, there is strong overlap by Jaccard index between the leading edges for the timepoints showing induced CREB1 target gene expression. By contrast, there is low overlap between blue nodes and red nodes indicating that the CREB1 target genes suppressed in the 4 timepoints are distinct from those induce at other timepoints.