AhR may be involved in Th17 cell differentiation in chronic hepatitis B

Abstract

AbstractTh17 cells which are crucial for host immunity have been demonstrated to increase HBV infection. However, the mechanism of the Th17 cell increase is unknown. Hence, the mechanism of Th17 cell enhancement is important to provide a theoretical foundation for chronic hepatitis B immunotherapy. This study included 15 instances in the healthy control (HC) and 15 cohorts in the chronic hepatitis B (CHB). Their CD4+T cells were isolated from their peripheral blood and then subjected to RNA transcriptome sequencing. Then, to identify target genes linked to Th17‐cell differentiation, DEGs associated with CHB were convergent with the Th17‐cell‐associated genes from the KEGG database. Hub genes of DEG and target genes linked to Th17 cells were analysed for correlation. The AhR‐related genes were located using the GeneMANIA database. To analyse the function of the genes, GO and KEGG pathways were employed. Protein–protein interaction network analysis employed the Metascape, STRING and Cytoscape databases. Finally, Western blotting and RT‐qPCR were used to validate AhR. A total of 348 differential genes were identified in CHB patients. CytoHubba was used for screening five hub genes associated with CHB: CXCL10, RACGAP1, TPX2, FN1 and GZMA. This study aimed to determine the mechanism of elevated Th17 cells in CHB. As a result, further investigation using the convergence of DGEs and Th17 cell‐related genes identified three target genes: AhR, HLA‐DQA1 and HLA‐DQB1, all of which were elevated in CHB. The three genes were primarily involved in immune response‐related processes, according to the GO enrichment analysis. Correlation analysis of CXCL10, RACGAP1, TPX2, FN1 and GZMA genes with AhR, HLA‐DQA1 and HLA‐DQB1 revealed that AhR was positively associated with CXCL10 and GZMA genes, which best respond to the severity of CHB disease. Combined with the role of AhR in Th17 cell differentiation, the genes AhR was chosen for confirmation by RT‐qPCR and WB in this study. The results showed that the CHB group had higher expression levels of AhR at both RT‐qPCR and WB levels. Furthermore, this study's findings revealed that AhR may contribute to the development of CHB by affecting the differentiation of Th17 cells.