RNA Sequencing Analyses and Validation of Immune-Related Genes in EBV-Specific Cytotoxic T Lymphocytes During Latent Infection

潜伏感染期间EBV特异性细胞毒性T淋巴细胞中免疫相关基因的RNA测序分析和验证

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Abstract

BACKGROUND: Epstein-Barr virus (EBV) infects more than 90% of the global population, but the immune mechanism underlying its infection remains incompletely understood. METHODS: In this study, transcriptomic profiling and validation of differentially expressed genes (DEGs) were performed between Epstein-Barr virus-specific cytotoxic T lymphocytes (EBV-CTLs) and nonspecific CTLs via RNA sequencing (RNA-seq). Bioinformatics analyses, including Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, functional molecular module construction, and key gene analysis, were conducted to explore the biological functions and potential regulatory mechanisms of the DEGs. RESULTS: A total of 1236 DEGs were identified in the EBV-CTL group compared with the nonspecific CTL group, including 645 upregulated genes and 591 downregulated genes. GO enrichment analysis revealed that these DEGs were localized mainly to cell membranes and MHC class II protein complexes and were involved in biological processes such as cellular defense, leukocyte activation, proliferation, differentiation, and chemotaxis. KEGG pathway enrichment analysis revealed that the JNK/p38 MAPK pathway was the most significantly enriched signaling pathway, with key DEGs including p38, HSP72, and components of the AP-1 transcription factor complex (mainly JUN and FOS). The functional molecular module construction revealed that the top-scoring modules were associated primarily with signal transduction, the inflammatory response, the immune response, and molecular interactions (eg, protein and receptor binding). Key gene analysis identified JUN, FOS, TNF, and STAT1 as potential hub genes involved in the EBV-specific immune response. CONCLUSION: Our transcriptomic analysis reveals the unique gene expression profile of EBV-CTLs and identifies the JNK/p38 MAPK pathway and hub genes (JUN, FOS, TNF, and STAT1) as critical regulators of the EBV-specific immune response. These findings provide novel insights into the molecular mechanism underlying EBV-specific immunity and potential targets for related therapeutic intervention.

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