Fig. 7: AMP-CpG immunization stimulates dynamic changes in immune gene transcript levels in NHP draining lymph nodes.

Rhesus macaques (n = 3) were immunized at week 0, 4 and 10 with 140 µg WH-01 RBD admixed with 5 mg of AMP-CpG. Lymph node FNAs were collected at baseline and 24 h post-week 10 boost. Nanostring analysis, using the NanoString nCounter® NHP Immunology Panel, was performed comparing gene expression in lymph node FNA between baseline and 24 h post boost. a Volcano plot representation of log-transformed transcript values of 770 genes at 24 h. Dotted horizontal line represents significance threshold of p = 0.05 (determined with Rosalind software); vertical dotted lines represent fold-change limits of ±1.5-fold change. b Heat map of gene transcripts that were differentially expressed. Shown are mock-subtracted, average Z-scores of gene transcript levels significantly (p < 0.05, as determined with Rosalind software) downregulated (≥−1.5-fold change, blue) or upregulated (≥1.5-fold change, red) relative to baseline values. Insignificant values with p ≥ 0.05 are shown in gray. Genes were clustered into 9 groups (box insert) using Gene Ontology databases and annotated at the top of the heat map: (1) adaptive immunity, (2) antigen processing and presentation, (3) APC activation, (4) apoptosis, (5) inflammation, (6) innate immunity, (7) interferon signaling, (8) migration, and (9) T cell anergy and exhaustion. c Fold-change bar graphs for selected gene transcripts. Statistical analysis was performed using Rosalind software.