Figure 6
DDX17 gene expression is downregulated post vaccination with gH1-Qbeta and WGCNA/BTM analysis reveals clusters of genes associated with downregulation of DDX17 and upregulation of HAI and CD4 T-cell proliferative responses. (A) Volcano plot comparing mRNA microarray data in whole blood samples from subjects at the pre-vaccination timepoint and at 21 days after administration of vaccine [top plots; Pre vs Post 1] and comparing pre-vaccination to 42 days after administration of the vaccine [bottom plots; Pre vs Post 2]. Subjects vaccinated with adjuvanted (left column plots) and non-adjuvanted (right column plots) gH1-Qbeta are depicted as labelled. (B) Normalized DDX17 mRNA levels from microarray analysis of whole blood from subjects vaccinated with adjuvanted or non-adjuvanted gH1-Qbeta at pre-vaccination and 21 days following the first (Post 1) and second (Post 2) vaccination. Horizontal bars represent the median value. Friedman test. ****P < 0.0001; ***P < 0.001; *P < 0.05 by Dunn’s test. ns, not significant. (C) PBMCs from the cohort vaccinated with non-adjuvanted gH1-Qbeta were analysed by flow cytometry for DDX17 protein expression in a number of identified immune cell populations. Horizontal bars represent the median value. Wilcoxon test. **P < 0.01; *P < 0.05; ns, not significant. (D) WGCNA analysis clustered genes with similar expression patterns into modules with randomly assigned colors. Each module was tested for correlation to traits of interest. Pearson correlation coefficients are stated in each box, with p values in brackets. Labels “Pre”, “Post1” and “Post2”, refer to Day 0, Day 21 and Day 42 post primary vaccination respectively.
