Enrichment analyses of the signal pathway.
We also performed the analysis on pathway enrichment of DE genes in 100μg ag85a/b DNA IM group and 50μg ag85a/b DNA EP group. The pathway analysis results revealed the action pathway ofag85a/b DNA vaccine (supplementary Table 5-8): (1) There were respectively 15 identical pathways in both up-regulated and down-regulated DE genes of the 100μg DNA IM group and 50μg DNA EP group. (2) The down-regulated pathways in the 100μg DNA IM group and 50μg DNA EP group were mainly related to nutrient digestion and absorption, hormone and neurotransmitter secretion, endocrine and neuroendocrine, for example, pancreatic secretion, protein digestion and absorption, maturity-onset diabetes of the young, insulin secretion, fat digestion and absorption, renin-angiotensin system, neuroactive ligand-receptor interaction, etc; the up-regulated pathways were mainly related to immune defense, substance metabolism and tumor, for example, ECM-receptor interaction, focal adhesion, protein digestion and absorption, PI3K-Akt signaling pathway, proteoglycans in cancer, etc.
After further analysis of the specific pathways, we found that the pancreatic secretion pathway down-regulated and Rap1 signal pathway up-regulated had particularly significant changes during the immunotherapy of the ag85a/b DNA vaccine on the mouse TB model. The former was closely related to the digestion and absorption of nutrients, and the latter was related to immunity. Therefore, we focused on the regulatory changes of these two pathways before and afterag85a/b DNA vaccine treatment to explore the therapeutic effect of ag85a/b DNA vaccine on damage recovery caused by MTB infection and the mechanism of immunotherapy (shown in Figure 4). We found that the expression levels of 15 DE genes in the pancreatic secretion pathway were significantly changed before and after ag85a/b DNA treatment, in which the expression levels of 12 DE genes, NBC1, GS, PRSS, CTRB1, CELA, CPA, CPB, PNLIP, PLRP1, CEL, PLRP2, and PLA2, increased significantly after MTB infection, but decreased significantly after ag85a/b DNA treatment. The expression levels of 15 DE genes in the Rap1 signal pathway were significantly changed before and afterag85a/b DNA treatment, in which the expression levels of 12 DE genes, GF, NMDAR, RTK, GPCR, ADAP, M-ras, Rap1, DOCK4, Epac, Talin, Profillin, p38MAPK, decreased significantly after MTB infection, but increased significantly after ag85a/b DNA treatment.