Background: Hepatitis B virus (HBV) is clinically associated with poor prognosis in diffuse large B-cell lymphoma (DLBCL), while cellular communication in the tumour microenvironment (TME) is recognized as a critical driver of tumour progression. Nevertheless, whether HBV infection mediates DLBCL cell-immune cell crosstalk remains undefined, with the precise mechanisms and associated key molecules remaining elusive.
Methods: SsGSEA, Cox regression (univariate/multivariate), WGCNA, and Kaplan–Meier analyses identified prognostic immune subsets and miRNAs in HBV+ DLBCL. Dual luciferase assay, qRT-PCR, western blot, ChIP, Co-IP, flow cytometry, enzyme-linked immunosorbent assay, immunohistochemistry, and murine models were employed together to evaluate CD4+ T cell dysfunction in vitro and in vivo. ScRNA-seq analyses encompassed clustering, pseudotemporal trajectory, and ligand–receptor networks to decode TME dynamics.
Results: TME profiling identified diminished CD4⁺ T cell infiltration as an independent predictor of poor survival in HBV⁺ DLBCL. Mechanistically, HBx-mediated down-regulation of miR-19a-3p activated the BAMBI/Wnt signalling pathway, thereby enhancing TGF-β1 secretion and suppressing the anti-tumour activity of CD4+ T cells. Single-cell analysis revealed that BAMBIhigh DLBCL cells engage CD4+ T cells via TGFB1-TGFBR2 pair, with TGFBR2 enriched in exhausted subsets of CD4+ T cells and shaping their dysfunctional fate. Therapeutic restoration of miR-19a-3p or blockade of TGF-β reinforced the CD4⁺ T cell anti-tumour activity and restrained the progression of HBx-overexpressing DLBCL in vivo.
Conclusions: HBx promoted TGF-β1 hypersecretion via miR-19a-3p repression-mediated Wnt/β-catenin activation, directly driving CD4+ T cell depletion and functional exhaustion in DLBCL. Our work provided important insights into the immune determinants of poor prognosis in HBV+ DLBCL, highlighting the pivotal role of CD4+ T cell dysfunction in driving disease progression and adverse clinical outcomes.
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2026 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.
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