AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level

IntroductionUnresolved hepatitis B virus (HBV) infection leads to a progressive state of HBV-specific immune dysfunctionality that characterizes chronic infection. The immune-competent adeno associated virus (AAV)-HBV mouse model is commonly used preclinically, though a comprehensive characterizatio...

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Main Authors: Nádia Conceição-Neto, Wim Pierson, Qinglin Han, Zhiyuan Yao, Qun Wu, Koen Dockx, Liese Aerts, Dries De Maeyer, Matthias Beyens, Koen Van den Berge, Chris Li, George Kukolj, Ren Zhu, Vinod Krishna, Ondřej Podlaha, Isabel Nájera, Ellen Van Gulck
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1421712/full
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author Nádia Conceição-Neto
Wim Pierson
Qinglin Han
Zhiyuan Yao
Qun Wu
Koen Dockx
Liese Aerts
Dries De Maeyer
Matthias Beyens
Koen Van den Berge
Chris Li
George Kukolj
Ren Zhu
Vinod Krishna
Ondřej Podlaha
Isabel Nájera
Ellen Van Gulck
author_facet Nádia Conceição-Neto
Wim Pierson
Qinglin Han
Zhiyuan Yao
Qun Wu
Koen Dockx
Liese Aerts
Dries De Maeyer
Matthias Beyens
Koen Van den Berge
Chris Li
George Kukolj
Ren Zhu
Vinod Krishna
Ondřej Podlaha
Isabel Nájera
Ellen Van Gulck
author_sort Nádia Conceição-Neto
collection DOAJ
description IntroductionUnresolved hepatitis B virus (HBV) infection leads to a progressive state of HBV-specific immune dysfunctionality that characterizes chronic infection. The immune-competent adeno associated virus (AAV)-HBV mouse model is commonly used preclinically, though a comprehensive characterization of the liver immune microenvironment and its translatability to human infection is still lacking. We investigated the intrahepatic immune profile of the AAV-HBV mouse model at a single-cell level and compared with data from CHB patients in immune tolerant (IT) and immune active (IA) clinical stages.MethodsImmune exhaustion was profiled through an iterative subclustering approach for cell-typing analyses of single-cell RNA-sequencing data in CHB donors and compared to the AAV-HBV mouse model 4-weeks and 24-weeks post-transduction to assess its translatability. This was confirmed using an exhaustion flow cytometry panel at 4 and 42-weeks post-transduction.ResultsUsing single-cell RNA-sequencing, CD8 pre-exhausted T-cells with self-renewing capacity (TCF7+), and terminally exhausted CD8 T-cells (TCF7-) were detected in the AAV-HBV model. These terminally exhausted CD8 T-cells (expressing Pdcd1, Tox, Lag3, Tigit) were significantly enriched versus control mice and independently identified through flow cytometry. Importantly, comparison to CHB human data showed a similar exhausted CD8 T-cell population in IT and IA donors, but not in uninfected individuals. DiscussionLong term high titer AAV-HBV mouse liver transduction led to T-cell exhaustion, as evidenced by expression of conventional immune checkpoint markers at mRNA and protein levels. In both IT and IA donors, a similar CD8 exhausted T-cell population was identified, with increased frequency observed in IA donors. These data support the use of the AAV-HBV mouse model to study classical T-cell exhaustion in HBV infection and the effect of immune-based therapeutic interventions.
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spelling doaj-art-c050bb440e0d428cb5f34b9112d85d512025-08-20T02:07:38ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-06-011610.3389/fimmu.2025.14217121421712AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell levelNádia Conceição-Neto0Wim Pierson1Qinglin Han2Zhiyuan Yao3Qun Wu4Koen Dockx5Liese Aerts6Dries De Maeyer7Matthias Beyens8Koen Van den Berge9Chris Li10George Kukolj11Ren Zhu12Vinod Krishna13Ondřej Podlaha14Isabel Nájera15Ellen Van Gulck16ID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Beerse, BelgiumID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Beerse, BelgiumID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Shanghai, ChinaID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Brisbane, CA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Shanghai, ChinaCharles River Laboratories, Beerse, BelgiumID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Beerse, BelgiumDiscovery Therapeutics and Molecular Pharmacology, Johnson & Johnson Innovative Medicine, Beerse, BelgiumDiscovery Therapeutics and Molecular Pharmacology, Johnson & Johnson Innovative Medicine, Beerse, BelgiumStatistics and Decision Sciences, Johnson & Johnson Innovative Medicine, Beerse, BelgiumID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Brisbane, CA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Brisbane, CA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Shanghai, ChinaID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Spring House, PA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Brisbane, CA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Brisbane, CA, United StatesID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Beerse, BelgiumIntroductionUnresolved hepatitis B virus (HBV) infection leads to a progressive state of HBV-specific immune dysfunctionality that characterizes chronic infection. The immune-competent adeno associated virus (AAV)-HBV mouse model is commonly used preclinically, though a comprehensive characterization of the liver immune microenvironment and its translatability to human infection is still lacking. We investigated the intrahepatic immune profile of the AAV-HBV mouse model at a single-cell level and compared with data from CHB patients in immune tolerant (IT) and immune active (IA) clinical stages.MethodsImmune exhaustion was profiled through an iterative subclustering approach for cell-typing analyses of single-cell RNA-sequencing data in CHB donors and compared to the AAV-HBV mouse model 4-weeks and 24-weeks post-transduction to assess its translatability. This was confirmed using an exhaustion flow cytometry panel at 4 and 42-weeks post-transduction.ResultsUsing single-cell RNA-sequencing, CD8 pre-exhausted T-cells with self-renewing capacity (TCF7+), and terminally exhausted CD8 T-cells (TCF7-) were detected in the AAV-HBV model. These terminally exhausted CD8 T-cells (expressing Pdcd1, Tox, Lag3, Tigit) were significantly enriched versus control mice and independently identified through flow cytometry. Importantly, comparison to CHB human data showed a similar exhausted CD8 T-cell population in IT and IA donors, but not in uninfected individuals. DiscussionLong term high titer AAV-HBV mouse liver transduction led to T-cell exhaustion, as evidenced by expression of conventional immune checkpoint markers at mRNA and protein levels. In both IT and IA donors, a similar CD8 exhausted T-cell population was identified, with increased frequency observed in IA donors. These data support the use of the AAV-HBV mouse model to study classical T-cell exhaustion in HBV infection and the effect of immune-based therapeutic interventions.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1421712/fullhepatitis B virussingle-cell transcriptomeAAV-HBV-infected mouse modelT cell exhaustionchronic HBV
spellingShingle Nádia Conceição-Neto
Wim Pierson
Qinglin Han
Zhiyuan Yao
Qun Wu
Koen Dockx
Liese Aerts
Dries De Maeyer
Matthias Beyens
Koen Van den Berge
Chris Li
George Kukolj
Ren Zhu
Vinod Krishna
Ondřej Podlaha
Isabel Nájera
Ellen Van Gulck
AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
Frontiers in Immunology
hepatitis B virus
single-cell transcriptome
AAV-HBV-infected mouse model
T cell exhaustion
chronic HBV
title AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
title_full AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
title_fullStr AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
title_full_unstemmed AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
title_short AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level
title_sort aav hbv mouse model replicates the intrahepatic immune landscape of chronic hbv patients at single cell level
topic hepatitis B virus
single-cell transcriptome
AAV-HBV-infected mouse model
T cell exhaustion
chronic HBV
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1421712/full
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