Oral Microbiome Diversity Matters on Nucleos(t)ide Analogue Cessation in Chronic Hepatitis B

Mahin Ghorbani; Agne Kvedaraite; Khaled Al-Manei; Choon Boon Too; Susanne Cederberg; Asgeir Johannessen; Dag Henrik Reikvam; Davide Valentini; Christopher Maucourant; Niklas K Björkström; Soo Aleman; Margaret Sällberg Chen

doi:10.1093/infdis/jiaf591

Oral Microbiome Diversity Matters on Nucleos(t)ide Analogue Cessation in Chronic Hepatitis B

J Infect Dis. 2025 Dec 2:jiaf591. doi: 10.1093/infdis/jiaf591. Online ahead of print.

Authors

Mahin Ghorbani¹, Agne Kvedaraite¹, Khaled Al-Manei^{1

2}, Choon Boon Too¹, Susanne Cederberg³, Asgeir Johannessen^{4

5

6}, Dag Henrik Reikvam^{5

6}, Davide Valentini⁷, Christopher Maucourant⁸, Niklas K Björkström⁸, Soo Aleman^{8

3}, Margaret Sällberg Chen^{1

7}

Affiliations

¹ Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Huddinge, Sweden.
² Division of Endodontics, Department of Restorative Dental Science, College of Dentistry, King Saud University, Riyadh 11545, Saudi Arabia.
³ Department of Infectious Diseases, Karolinska University Hospital, Stockholm, Sweden.
⁴ Department of Infectious Diseases, Vestfold Hospital Trust, Tønsberg, Norway.
⁵ Department of Infectious Diseases, Regional Advisory Unit for Imported and Tropical Diseases, Oslo University Hospital, Oslo, Norway.
⁶ Faculty of Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁷ Department of Cellular Therapy and Allogeneic Stem Cell Transplantation (CAST), Karolinska University Hospital Huddinge and Karolinska Comprehensive Cancer Center, Stockholm, Sweden.
⁸ Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Karolinska University Hospital, Huddinge, Sweden.

PMID: 41328917
DOI: 10.1093/infdis/jiaf591

Abstract

Background: Withdrawal of nucleos(t)ide analog (NUC) therapy in HBeAg-negative chronic hepatitis B (CHB) may lead to functional cure in a subset of patients. Although gut microbiota is known to influence both CHB-progression and treatment outcomes, the oral microbiome in NUC-cessation remains unexplored.

Methods: This long-term longitudinal study aimed to explore the oral microbiome in CHB patients who have been on NUC therapy >2 years having a planned NUC cessation. Oral microbiome composition was analyzed in 110 saliva samples across seven timepoints from 18 HBeAg-negative patients with 36 months of follow-up. Favorable outcome was defined as either HBsAg loss or decline of >1 log10 or sustained off-therapy HBV DNA level <2000 IU/ml during year 3. Hepatic flare was defined as ALT >80 U/L or 2x baseline level.

Results: The overall microbial composition remained stable during the study period. Patients with favorable outcome (n=8) showed consistently higher alpha diversities (Shannon and Simpson indices, p<0.001) from baseline, with lower inter-sample variations across all timepoints (Bray-Curtis and Jaccard distances, p<0.05), compared to unfavorable (n=10). HBsAg, ALT and AST correlated inversely with several Prevotella taxa with specific pathways (Spearman´s rho>-0.5, p<0.01). Unfavorable outcome and high HBsAg level correlated with opportunistic taxa Haemophilus parainfluenzae and Porphyromonas catoniae at multiple timepoints. Random forest model incorporating ANCOM-validated microbial markers predicting favorable vs unfavorable outcome achieved higher predictive performance than clinical markers alone (AUC 0.78 vs. 0.66).

Conclusion: Our exploratory study suggests that oral microbiome profiling at NUC cessation in HBeAg-negative CHB could support prognostication of virological outcome.

Keywords: HBeAg; HBsAg; antiviral treatment; cure; hepatic flare; nucleos(t)ide analog discontinuation; oral microbiome; viral hepatitis.