Transcriptional analysis of human peripheral blood mononuclear cells stimulated by Mycobacterium tuberculosis antigen

• DOI: 10.3389/fcimb.2023.1255905
• Publication Date: 2023-09-26T06:01:06Z
• Authors: Jing Wei; Fangzheng Guo; Yamin Song; Kun Xu; Feiyang Lin; Kangsheng Li; Baiqing Li; Zhongqing Qian; Xiaojing Wang; Hongtao Wang; Tao Xu
• Keywords: KEGG

Background Mycobacterium tuberculosis antigen (Mtb-Ag) is a polypeptide component with molecular weight of 10-14 kDa that obtained from the supernatant H37Ra strain after heat treatment. It stimulates activation and proliferation γδT cells in blood to produce an immune response against tuberculosis. Mtb-Ag therefore crucial for classifying detecting central genes key pathways involved TB initiation progression. Methods In this study, we performed high-throughput RNA sequencing peripheral mononuclear (PBMC) Mtb-Ag-stimulated control samples identify differentially expressed used them gene ontology (GO) Kyoto Encyclopedia Genomes (KEGG) enrichment analysis. Meanwhile, PPI protein interaction network Cytoscape analysis qRT-PCR verify differential expression. Single-gene (GSEA) was further elucidate potential biological functions genes. Analysis cell infiltration correlation using R language. Results We identified 597 stimulated PBMCs. KEGG GSEA analyzed cellular related function, DEGs were found be primarily TNF signaling pathway, IL-17 JAK-STAT cytokine-cytokine receptor interactions, NF-κB pathway. Wayne GSEA, KEGG, protein-protein (PPI) showed 34 genes, including PTGS2, IL-1β, IL-6, IFN-γ et al., co-expressed five all up-regulated by stimulation. Twenty-four qRT-PCR, fourteen (SERPINB7, IL20, IFNG, CSF2, TNF-α, IL36G, IL6, IL10, IL1A, CXCL1, CXCL8, IL4, CXCL3) ten down-regulated (RTN1, CSF1R CD14, C5AR1, CXCL16, PLXNB2, OLIG1, EEPD1, ENG, CCR1). These findings consistent RNA-Seq results. Conclusion The transcriptomic features associated provide scientific basis exploring intracellular mechanisms Mtb. However, more studies on these stimulation are needed underlying pathologic during Mtb infection.