Identifying the Key Genes of Epstein Barr Virus Regulated Tumour Immune Microenvironment of Diffuse Large B Cell Lymphoma

Objective: Epstein Barr virus (EBV) plays a crucial role in the pathogenesis of diffuse large B cell lymphoma (DLBCL) by infecting normal cells and influencing the composition of the tumor immue microenvironment (TIME). However, a systematic bioinformatics analysis of key genes regulating immune infiltration in EBV+ DLBCL is currently lacking.

Methods: We utilized the GEO database to identify differential genes (DEGs) and analyze immune infiltration profiles in EBV+ DLBCL (N=23) and EBV- DLBCL (N=28). Weighted gene co-expression analysis (WGCNA) was employed to identify gene modules associated with EBV-induced immune infiltration. Immune-related genes were sourced from the ImmPort database. Immune-related hub genes in EBV+ DLBCL were identified through an integrated analysis of DEGs, WGCNA, ImmPort data, and protein-protein interaction (PPI) networks. Additionally, single-cell RNA sequencing (scRNA-seq) was performed on tumor tissue from one EBV+ DLBCL case to validate hub gene expression in immune cells.

Results: We identified 1215 genes with differential expression between EBV+ and EBV- DLBCL, and enrichment analysis showed that DEGs were mainly enriched in immune-related pathways. EBV+ DLBCL exhibited distinct profiles of tumor-infiltrating immune cells compared to EBV- DLBCL, with more plasma cells, CD8+ T cells, T cell CD4 memory-activated, NK cell-activated, monocytes, macrophage M1, dendritic cell-activated, thymocyte-activated, oocytes, neutrophils and less B-cell naive, T-cell CD4 naive, T-cell CD4 memory resting, T-cell follicular helper, T-cell regulatory, macrophage M0, and dendritic cell resting. Ultimately, we identified three immune-related hub genes-CD28, IL18, and CD247-that were down-regulated and correlated with poor prognosis in EBV+ DLBCL. scRNA-seq confirmed that these hub genes were predominantly expressed in T cell, NK cell and monocytes clusters.

Conclusions: EBV infection significantly alters the TIME of DLBCL, potentially through the down-regulation of CD28, IL18, and CD247, which may contribute to the observed poor prognosis in EBV+ DLBCL patients. These identified hub genes provide new insights into potential prognostic markers and therapeutic targets for EBV+ DLBCL.

No relevant conflicts of interest to declare.