Co-expression network analysis identifies innate immune signatures for Albizia julibrissin saponin active fraction-adjuvanted avian influenza vaccine

Albizia julibrissin saponin active fraction (AJSAF) is a promising adjuvant candidate, but its innate immune response mechanisms remain unclear. Here, the quadriceps muscles from the mice injected intramuscularly with AJSAF alone or in combination with ovalbumin and avian influenza vaccine (rL-H5) were subjected to gene microarray. Antigen- and AJSAF-related modules with intramodular hub genes were identified and functionally analyzed using weighted gene co-expression network analysis (WGCNA) and gene set enrichment analysis (GSEA). AJSAF induced early innate immune responses at the injection site, characterized by cytokine production and neutrophil recruitment. AJSAF mainly elicited the expression of "Th1 immune response" and "Neutrophils" genes such as CCL2, CXCL1, CXCL5, IL-1β, IL-6, IL-33, S100A8, and S100A9, whereas these two gene sets were negatively enriched for rL-H5. AJSAF-specific long noncoding RNAs MIRT1 and MIRT2 could function as inflammatory mediators, whereas function unknown TINCR was co-expressed with immune response genes including CCL2, CCL4, CCL7, CSF3, CXCL5, IL-33, S100A8, and S100A9. Finally, the innate immune molecular mechanisms of adjuvant action of AJSAF and the potential signatures were proposed. These findings expanded the current knowledge on the mechanisms of action of saponin-based adjuvants.