Jiming Zhang

The funder National Key Research and Development Program of China (Grant no. 2023YFC2308500) was erroneously omitted from the Funding statement. The correct Funding statement reads:"The author(s) declared that financial support was received for this work and/or its publication. The original version of this article has been updated.

Introduction: SARS-CoV-2/HBV coinfections are increasingly common as SARS-CoV-2 continues to evolve. The interferon (IFN) therapy for HBV infection might also have antiviral effects against SARS-CoV-2. To understand the effects of IFN on SARS-CoV-2 infection, we compared the clinical characteristics between IFN-treated chronic hepatitis B (CHB) patients and nucleo(s)tide analogs (NAs)-treated CHB patients during their SARS-CoV-2 infection. Methods: Our cohort study enrolled patients with primary SARS-CoV-2 infections among IFN-treated and NAs-treated CHB patients during the Omicron wave in Shanghai, China, from December 7 2022 to January 23 2023. Clinical characteristics and self-reported acute symptoms during SARS-CoV-2 infection were collected by telephone questionnaire in January 2023. Results: The four most common symptoms were fever, fatigue, cough and sore throat among all enrolled participants, regardless of their treatment. Interestingly, Pegylated interferon-alpha (Peg-IFNα) treated CHB patients had a significantly higher proportion of asymptomatic infection than NAs-treated CHB patients (14.53% vs. 2.70%, p = 0.002). Additionally, IFN-treated CHB patients had significantly lower duration of fever ( p = 0.018), less usage of antipyretics ( p < 0.0001), less occurrence of muscle and/or joint pain ( p < 0.0001), less occurrence of headache ( p < 0.0001) and less occurrence of runny nose ( p < 0.0001) compared to NAs-treated CHB patients. Conclusion: In this real-world cohort of CHB patients infected with SARS-CoV-2 during the Omicron wave, Peg-IFNα treatment was associated with milder self-reported acute clinical manifestations, including a higher proportion of asymptomatic infection and reduced occurrence of several common symptoms.

Background: The preS1 region of the HBV large surface protein binds sodium taurocholate cotransporting polypeptide (NTCP) to mediate viral entry, but the serological and functional relevance of circulating preS1 antigen and anti-preS1 antibodies in chronic hepatitis B (CHB) is not well defined. Objective: To characterise serum preS1 antigen and anti-preS1 IgG across CHB and evaluate their virological and functional significance. Design: ELISAs for preS1 antigen and anti-preS1 IgG were developed and applied to 549 individuals with chronic HBV infection, 107 individuals with functional cureand 110 vaccinated healthy controls. Neutralising activity was assessed using HBV and hepatitis D virus (HDV) infection models, preS1-NTCP competitive assays and peptide microarrays. Baseline anti-preS1 IgG was evaluated in 51 patients discontinuing nucleos(t)ide analogue (NUC) therapy. Results: PreS1 antigen was highest in hepatitis B e antigen (HBeAg)-positive patients and correlated with HBV DNA, hepatitis B surface antigen and HBeAg. Anti-preS1 IgG was detectable across disease phases, inversely associated with HBV DNA, positively correlated with alanine aminotransferase/aspartate aminotransferase and remained detectable in some individuals with functional cure. Sera with low preS1 and high anti-preS1 IgG showed potent HBV/HDV neutralising activity by inhibiting viral entry and blocking preS1-NTCP binding. Dominant linear epitopes mapped to preS1 regions involved in NTCP binding. High baseline anti-preS1 IgG was associated with sustained virological suppression after NUC withdrawal. Conclusion: PreS1 antigenaemia and anti-preS1 IgG exhibit distinct serological patterns linked to HBV replication and endogenous viral control. High anti-preS1 IgG confers potential neutralising activity and associates with sustained off-therapy suppression, highlighting its functional relevance as an indicator of host viral control.

Chronic hepatitis B virus (HBV) infection (CHB) progresses through distinct clinical phases characterized by dynamic immune responses and changes in intrahepatic gene expression. While hepatic transcriptomes have been studied, hepatic microRNA (miRNA) profiles across the natural history of CHB remain poorly defined. In this study, we aimed to characterize hepatic miRNA expression and its association with liver cell composition across CHB phases. Liver biopsies from 43 treatment-naïve CHB patients across four clinical phases, immune tolerance (IT), immune active (IA), inactive carrier (IC), and HBeAg-negative hepatitis (ENH), were analyzed using miRNA microarrays and mRNA sequencing. Hepatic cell composition was inferred using CIBERSORT and expression of 92 cell-type marker genes. Differentially expressed miRNAs were identified by Mann–Whitney U test, and clustering and correlation analyses were performed to assess associations with immune and liver cell markers. Consistent with previous studies, immune cell infiltration varied by phase, peaking in IA and ENH phases. Sixty-five miRNAs were significantly differentially expressed (|log₂FC| > 0.585), including immune-associated miRNAs such as miR-155-5p, miR-150-5p, and miR-142-5p. Twenty-two miRNAs elevated in IA phase were associated with macrophage and T cell markers. Hepatocyte-enriched miRNAs (e.g., miR-122-5p, miR-192-5p) remained highly expressed across all phases. Clustering analysis identified five miRNA expression patterns, with immune-associated clusters upregulated in IA and ENH phases. Correlation analysis confirmed strong associations between immune-phase miRNAs and αβ T cell markers. Overall, these findings indicate that hepatic miRNA profiles are largely stable across CHB phases, with immune cell infiltration associated with selective miRNA changes, particularly in IA and ENH phases.

Chronic hepatitis B (CHB) remains incurable due to the immune system's tolerance toward the hepatitis B virus (HBV) surface antigen (HBsAg). This study aimed to achieve a functional cure by breaking HBV tolerance through immunotherapy. CHB patients were treated with either standard nucleotide analog (NA) therapy (Adefovir Dipivoxil, ADV) (Cohort 1) or ADV combined with interferon‐alpha (IFN‐α) (Cohort 2). Additionally, a third cohort received the THRIL‐GM‐Vac regimen: three low‐dose GM‐CSF injections followed by one dose of the HBV vaccine, alongside standard treatment. THRIL‐GM‐Vac treatment (Cohort 3) achieved a significant 2log10 reduction in HBsAg levels in 21.7% of participants, and 8.7% HBsAg clearance in Cohort 3 compared to 0% and 4.17% in Cohorts 1 and 2, respectively. Furthermore, THRIL‐GM‐Vac significantly reduced HBV‐specific tolerogenic T cells (Tregs), explaining the sustained HBsAg decrease. Upregulation of anti‐HBV T cell responses confirmed THRIL‐GM‐Vac's ability to disrupt HBV tolerance and enhance HBsAg‐specific cellular immunity. This suggests its potential effectiveness in treating individuals with moderate to low HBsAg levels. THRIL‐GM‐Vac treatment in Cohort 3 resulted in 8.7% HBsAg clearance alongside Treg depletion and enhanced anti‐viral T cell responses. These findings present a promising strategy to overcome immunotolerance and potentially combat chronic HBV infection.

Chronic hepatitis B (CHB) is the leading cause of acute-on-chronic liver failure (ACLF) in China and other Asian countries. A defining immunopathological feature of hepatitis B virus-related ACLF (HBV-ACLF) is immune paralysis, which significantly increases susceptibility to secondary bacterial infections and contributes to poor clinical outcomes. A critical determinant of this immunosuppressed state is impaired antigen presentation due to reduced human leukocyte antigen DR (HLA-DR) expression on monocytes; however, the epigenetic mechanism underlying HLA-DR downregulation in HBV-ACLF remains unclear. Methyl-CpG-binding domain protein 2 (MBD2), an epigenetic reader of DNA methylation, has been implicated in the regulation of monocyte-macrophage function in inflammatory diseases, but its role in HBV-ACLF pathophysiology remains to be fully elucidated. In this study, bulk RNA sequencing (RNA-seq) of circulating monocytes from patients with HBV-ACLF showed a transcriptional profile consistent with immune paralysis, characterized by suppressed antigen presentation and inflammatory pathways, alongside pronounced activation of epigenetic regulatory programs. MBD2 expression was subsequently assessed using immunohistochemistry (IHC), reverse transcription quantitative PCR (RT-qPCR), and flow cytometry. Monocyte MBD2 expression was significantly upregulated in HBV-ACLF and was positively correlated with disease severity (r = 0.2797, P = 0.0182), systemic inflammation indices, and clinical prognosis. To delineate the mechanistic role of MBD2, an MBD2-knockout THP-1 cell model was established and subjected to integrated RNA-seq and assay for transposase-accessible chromatin sequencing (ATAC-seq) following differentiation and lipopolysaccharide (LPS) stimulation. The results showed that MBD2 deficiency significantly increased chromatin accessibility and transcriptional activation of genes involved in antigen presentation and pro-inflammatory responses, including pathways related to major histocompatibility complex (MHC) class II synthesis. Concurrently, enhanced promoter accessibility and activation of transcription factors associated with HLA-II class expression were observed, and increased surface HLA-DR expression was confirmed by flow cytometry. Collectively, these findings suggest that MBD2 epigenetically represses HLA-DR expression in monocytes, leading to impaired antigen presentation and immune paralysis, thereby predisposing patients with HBV-ACLF to secondary bacterial infections. Therefore, MBD2 may serve as a novel biomarker for disease progression and a potential therapeutic target for restoring immunological competence in patients with ACLF.

Acute‐on‐chronic liver failure (ACLF), particularly hepatitis B virus‐related ACLF (HBV‐ACLF), is a severe condition with high short‐term mortality. This study aimed to evaluate the prognostic value of circulating cell‐free mitochondrial DNA (cf‐mtDNA) in predicting short‐term mortality in HBV‐ACLF patients. A total of 320 HBV‐ACLF patients were included in the study population, with 192 patients in the derivation cohort and 128 in the validation cohort. Plasma cf‐mtDNA levels were quantified using qPCR. Plasma cf‐mtDNA levels were significantly elevated in HBV‐ACLF patients (3.95 log ₁₀ copies/μL) compared to healthy controls (2.99 log ₁₀ copies/μL) and patients with chronic liver disease (3.03 log ₁₀ copies/μL) ( p < 0.001). Plasma cf‐mtDNA levels progressively increased with disease severity and were associated with multi‐organ failure. Sequential measurements of cf‐mtDNA showed a significant decrease in survivors (4.00 vs. 3.78 log ₁₀ copies/μL, p = 0.019), while non‐survivors maintained persistently elevated levels with minimal change (4.19 vs. 4.15 log ₁₀ copies/μL, p = 0.359). The cut‐off value of 3.9 log ₁₀ copies/μL for cf‐mtDNA effectively stratified patients into high‐ and low‐risk groups, with significantly lower survival rates in the high‐risk group (28‐day: 46.2% vs. 86.0%, p < 0.001; 90‐day: 37.7% vs. 72.1%, p < 0.001). Using cf‐mtDNA in combination with key clinical parameters, we developed a novel prognostic score that exhibited superior predictive accuracy for 28‐ and 90‐day mortality (AUROCs: 0.907 and 0.906, respectively), outperforming established prognostic scores (all p < 0.05). These findings were validated in an independent cohort. Cf‐mtDNA is a superior biomarker for predicting short‐term mortality in HBV‐ACLF. Its association with multi‐organ failure and disease severity highlights the potential utility in early risk assessment and treatment optimization.

Patients with chronic hepatitis B virus (HBV) infection in the immune-tolerant phase may still experience hepatic inflammation and disease progression, and could benefit from early antiviral treatment. This study aimed to investigate changes in the cumulative hepatitis B surface antigen (HBsAg)/HBV DNA ratio in immune-tolerant patients during the transition to the immune-active phase, and to evaluate its potential in predicting the risk of disease progression.