RNA interference-mediated hepatitis B virus antigen shutdown enhances the antiviral immune effects of PEGylated interferon-α via T and B cell crosstalk remodeling
4 affiliations
- 1Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Research Unit of Cure of Chronic Hepatitis B Virus Infection (CAMS), Shanghai Frontiers Science Center of Pathogenic Microbes and Infection, School of Basic Medical Sciences, Shanghai Medical College Fudan University, Shanghai, China
- 2Shanghai Institute of Infectious Disease and Biosecurity, Shanghai, China
- 3Liver Cancer Institute, Zhongshan Hospital, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University, Shanghai, China
- 4National Key Laboratory of Immunity & Inflammation, Department of Pathogen Biology, Naval Medical University, Shanghai, China
Abstract
Background/Aims: PEGylated interferon-α (PEGIFNα) shows promise in treating chronic hepatitis B (CHB), yet patient response remains suboptimal. While suppressing hepatitis B virus (HBV) antigens by RNA interference (RNAi) could enhance PEGIFNα efficacy in CHB patients, the underlying immunological mechanisms remain obscure.
Methods: Using our newly established extracellular humanized IFNAR mouse model of chronic HBV infection, we evaluated the efficacy of a GalNac-conjugated siRNA alone or in combination with PEGIFNα. Phenotypic and functional characteristics of immune cells were assessed by flow cytometry, ELISpot, and single-cell RNA sequencing (scRNA-seq).
Results: High circulating HBsAg reduced antiviral effects and immune responsiveness of PEGIFNα. Combined PEGIFNα and RNAi therapy synergistically and durably suppressed HBsAg (~4log10 IU/mL, vs. PBS) and achieved HBsAg seroconversion in ~30% of mice, outperforming either monotherapy. Mechanistically, PEGIFNα enhanced global T and B cell function, whereas combined therapy further amplified HBV-specific T and B cell responses. scRNA-seq analysis indicated that combined therapy attenuated inhibitory B cell-B cell interactions, strengthened MHC-I-mediated T cell crosstalk, and enhanced MHC-II signaling networks across B cells and hepatocytes/CD8+ T cells, collectively improving the lymphocyte functionality and facilitating HBsAg seroconversion. Reinforcing MHC-I/II signaling with agonistic antibodies (α4-1BB, αCD40) or IL-2-Fc further potentiated antiviral immune responses of combinational regimen.
Conclusions: Combined RNAi and PEGIFNα therapy exerts synergistic antiviral effects by relieving HBV antigen-induced immune tolerance and orchestrating a functional T cell-B cell crosstalk network centered on MHC-I/II signaling. Enhancing antigen presentation pathways represents a promising adjunctive strategy to improve functional cure rate of CHB.
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BibTeX
@article{zai2026interferencemediated,
title = {{RNA interference-mediated hepatitis B virus antigen shutdown enhances the antiviral immune effects of PEGylated interferon-α via T and B cell crosstalk remodeling}},
author = {Wenjing Zai and Kongying Hu and Mengying He and Ziyang Song and Pan Gao and Chen Luo and Minxiang Xie and Asha Ashuo and Min Yang and Jieliang Chen and Zhenghong Yuan},
journal = {Clinical and Molecular Hepatology},
year = {2026},
month = sep,
doi = {10.3350/cmh.2026.0385},
url = {https://doi.org/10.3350/cmh.2026.0385},
note = {CC BY-NC 3.0},
}
RIS
TY - JOUR
TI - RNA interference-mediated hepatitis B virus antigen shutdown enhances the antiviral immune effects of PEGylated interferon-α via T and B cell crosstalk remodeling
AU - Wenjing Zai
AU - Kongying Hu
AU - Mengying He
AU - Ziyang Song
AU - Pan Gao
AU - Chen Luo
AU - Minxiang Xie
AU - Asha Ashuo
AU - Min Yang
AU - Jieliang Chen
AU - Zhenghong Yuan
PY - 2026
DA - 2026/09/11
JO - Clinical and Molecular Hepatology
DO - 10.3350/cmh.2026.0385
UR - https://doi.org/10.3350/cmh.2026.0385
AB - Background/Aims: PEGylated interferon-α (PEGIFNα) shows promise in treating chronic hepatitis B (CHB), yet patient response remains suboptimal. While suppressing hepatitis B virus (HBV) antigens by RNA interference (RNAi) could enhance PEGIFNα efficacy in CHB patients, the underlying immunological mechanisms remain obscure. Methods: Using our newly established extracellular humanized IFNAR mouse model of chronic HBV infection, we evaluated the efficacy of a GalNac-conjugated siRNA alone or in combination with PEGIFNα. Phenotypic and functional characteristics of immune cells were assessed by flow cytometry, ELISpot, and single-cell RNA sequencing (scRNA-seq). Results: High circulating HBsAg reduced antiviral effects and immune responsiveness of PEGIFNα. Combined PEGIFNα and RNAi therapy synergistically and durably suppressed HBsAg (~4log10 IU/mL, vs. PBS) and achieved HBsAg seroconversion in ~30% of mice, outperforming either monotherapy. Mechanistically, PEGIFNα enhanced global T and B cell function, whereas combined therapy further amplified HBV-specific T and B cell responses. scRNA-seq analysis indicated that combined therapy attenuated inhibitory B cell-B cell interactions, strengthened MHC-I-mediated T cell crosstalk, and enhanced MHC-II signaling networks across B cells and hepatocytes/CD8+ T cells, collectively improving the lymphocyte functionality and facilitating HBsAg seroconversion. Reinforcing MHC-I/II signaling with agonistic antibodies (α4-1BB, αCD40) or IL-2-Fc further potentiated antiviral immune responses of combinational regimen. Conclusions: Combined RNAi and PEGIFNα therapy exerts synergistic antiviral effects by relieving HBV antigen-induced immune tolerance and orchestrating a functional T cell-B cell crosstalk network centered on MHC-I/II signaling. Enhancing antigen presentation pathways represents a promising adjunctive strategy to improve functional cure rate of CHB.
SN - 2287-2728
SN - 2287-285X
ER -