Conceição-Neto Nádia, Pierson Wim, Han Qinglin, Yao Zhiyuan, Wu Qun, Dockx Koen, Aerts Liese, De Maeyer Dries, Beyens Matthias, den Berge Koen Van, Li Chris, Kukolj George, Zhu Ren, Krishna Vinod, Podlaha Ondřej, Nájera Isabel, Van Gulck Ellen
ID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Beerse, Belgium.
ID Discovery, Infectious Diseases Therapeutic Area, Johnson & Johnson Innovative Medicine, Shanghai, China.
Front Immunol. 2025 Jun 18;16:1421712. doi: 10.3389/fimmu.2025.1421712. eCollection 2025.
Unresolved 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.
Immune 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.
Using single-cell RNA-sequencing, CD8 pre-exhausted T-cells with self-renewing capacity (), and terminally exhausted CD8 T-cells () were detected in the AAV-HBV model. These terminally exhausted CD8 T-cells (expressing ) 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.
Long 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.
未解决的乙型肝炎病毒（HBV）感染会导致一种以慢性感染为特征的HBV特异性免疫功能障碍的进行性状态。具有免疫活性的腺相关病毒（AAV）-HBV小鼠模型通常在临床前使用，尽管仍缺乏对肝脏免疫微环境及其与人类感染的可转化性的全面表征。我们在单细胞水平上研究了AAV-HBV小鼠模型的肝内免疫谱，并与免疫耐受（IT）和免疫活跃（IA）临床阶段的慢性乙型肝炎（CHB）患者的数据进行了比较。
通过迭代亚聚类方法对CHB供体的单细胞RNA测序数据进行细胞分型分析，以分析免疫耗竭情况，并与转导后4周和24周的AAV-HBV小鼠模型进行比较，以评估其可转化性。在转导后4周和42周使用耗竭流式细胞术面板对此进行了确认。
使用单细胞RNA测序，在AAV-HBV模型中检测到具有自我更新能力的CD8预耗竭T细胞（ ）和终末耗竭的CD8 T细胞（ ）。与对照小鼠相比，这些终末耗竭的CD8 T细胞（表达 ）显著富集，并通过流式细胞术独立鉴定。重要的是，与CHB人类数据的比较显示，IT和IA供体中存在类似的耗竭CD8 T细胞群体，但未感染个体中没有。
长期高滴度AAV-HBV小鼠肝脏转导导致T细胞耗竭，这在mRNA和蛋白质水平上通过传统免疫检查点标志物的表达得到证明。在IT和IA供体中，均鉴定出类似的CD8耗竭T细胞群体，在IA供体中观察到频率增加。这些数据支持使用AAV-HBV小鼠模型来研究HBV感染中的经典T细胞耗竭以及基于免疫的治疗干预的效果。
