AAV8-1.3HBV Mouse Model

Model Description

Hepatitis B virus (HBV) exhibits strict species specificity, limiting preclinical studies to non-human primates like chimpanzees, which face ethical and logistical constraints. Our AAV8-1.3HBV mouse model overcomes these challenges by delivering the full HBV genome via recombinant adeno-associated virus (rAAV8), enabling:

• High seroconversion rates: 88% HBsAg positivity within 4 weeks
• Sustained viremia: Detectable HBsAg/HBeAg for ≥24 weeks
• Humanized pathology: Liver inflammation and fibrosis progression

Key Advantages vs Traditional Models:
✓ Cost-effective scalability vs primate models
✓ Controlled viral replication (1.3x HBV genome coverage)
✓ Patent-protected transgenic lines for comparative studies

Applications

• Antiviral drug screening (NAs, siRNA, capsid inhibitors)
• HBV vaccine efficacy testing
• Viral persistence mechanism studies
• Immune modulator development (TLR agonists, checkpoint inhibitors)

Modeling Protocol —— rAAV8-1.3HBV Tail Vein Injection

1. Vector Preparation:

• rAAV8-1.3HBV construct (1.3x overlength HBV genome, genotype D)
• Viral titer: 1×10¹² vg/mL

2. Procedure:

• Inject 100 μL vector via tail vein (1×10¹¹ vg/mouse)
• Monitor HBsAg seroconversion weekly (ELISA)

3. Modeling Timeline:

• Week 4: Peak HBsAg expression (≥500 IU/mL)
• Week 12-24: Chronic phase with liver fibrosis (METAVIR ≥F2)
  

  Fig.1 Construction of rAAV-1.3 HBV model 

  Validation & Testing

  Category	Parameters
  Virological Analysis	• Serum HBsAg/HBeAg ELISA ∙ HBV DNA qPCR (log<sub>10</sub> IU/mL)
  Liver Function	ALT/AST levels ∙ Albumin/globulin ratio
  Histopathology	H&E staining ∙ Sirius Red collagen quantification ∙ HBsAg IHC
  Immune Profiling	Flow cytometry: CD8+ T cells ∙ PD-1/TIM-3 exhaustion markers
  Molecular Analysis	cccDNA quantification (Southern blot) ∙ HBV RNA RT-PCR

   

  Success Criteria
  ✓ Seroconversion: HBsAg >5 IU/mL by Week 4
  ✓ Viral persistence: HBV DNA >10⁴ IU/mL at Week 12
  ✓ Fibrosis progression: Collagen area ≥5% (Sirius Red)

  ---

   

  Technical Advantages

  Feature	Our Model	Traditional Models
  Infection Rate	88% ± 5%	≤50% (hydrodynamic injection)
  Chronicity	24+ weeks	8-12 weeks
  Clinical Relevance	Genotype D (global dominant)	Limited genotype coverage

Data