ArticleClinical and experimental vaccine research2025
Enhanced replication of a hepatitis A virus vaccine strain via adaptation in Vero cells.
Article in Clinical and experimental vaccine research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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7 authors.
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Abstract
Purpose: Hepatitis A virus (HAV) production has been limited by its slow replication rate and reliance on diploid cell lines like MRC-5, which present challenges in scalability, passage limitations, and serum-free culture conditions. This study aimed to develop an HAV vaccine strain with enhanced replication capacity. Materials and Methods: We generated a reverse genetically modified HAV vaccine strain (RG-HAV) and adapted it to Vero cells through sequential culturing. Replication rates of RG-HAV and a commercially used strain, HM-175, were compared in Vero and MRC-5 cells. Nucleotide sequences, including coding and non-coding regions like the internal ribosomal entry site (IRES), were analyzed. Structural assessments included 3-dimensional modeling of IRES and relative codon deoptimization analysis of the capsid. Immunogenicity was evaluated by measuring HAV-specific antibody responses in mice. Results: Vero-adapted RG-HAV achieved a 30-fold increase in production yield compared to initial transfection. In Vero cells, RG-HAV peaked at 15 days post-infection, compared to 20 days for HM-175. In MRC-5 cells, RG-HAV and HM-175 reached peak production at 10 and 15 days, respectively. RG-HAV produced over 5-fold more HAV in Vero cells and 8-fold more in MRC-5 cells than HM-175. Sequence analysis revealed nine amino acid differences in RG-HAV structural proteins and five nucleotide changes in the type III IRES region, potentially enhancing IRES functionality. Immunization with inactivated RG-HAV with alum hydroxide induced HAV-specific antibody responses in mice. Conclusion: RG-HAV offers enhanced replication and production yields, supporting its potential in advancing HAV vaccine development.
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