Nanoparticles targeting liver sinusoidal endothelial cells improve AAV gene transfer and promote tolerance-spreading to capsid and transgene epitopes
Nanoparticles targeting liver sinusoidal endothelial cells improve AAV gene transfer and promote tolerance-spreading to capsid and transgene epitopes
Presented by Sabine Fleischer at the 19th International Congress on Immunology Vienna 2025
Authors:
Summary
Introduction: Adeno-associated viral (AAV) gene transfer is still impeded by deleterious immune responses against capsid- and transgene-derived antigens. We developed an innovative approach for inducing antigen-specific tolerance to tackle these limitations. For that, nanoparticle-carriers were designed to preferentially deliver antigenic peptides to liver sinusoidal endothelial cells (LSECs) exploiting their natural potential of cross-presenting antigens and tolerizing T cell responses.
Objectives: Different mouse models of AAV-mediated gene transfer to muscle and liver were used to assess whether nanoparticles coupled to capsid- and transgene-derived peptides can effectively inhibit vector- and transgene-specific immune responses and improve transgene persistence.
Methods: Nanoparticle-peptide-carriers were intravenously administered at predefined timepoints in the context of muscle- and liver-directed AAV-gene transfer. T- and B-cell responses were analysed by ELISpot, flow cytometry, and ELISA; transgene expression by qPCR. Non-parametric Mann-Whitney test and one-way ANOVA were used to compare differences between two or multiple groups, respectively.
Results: LSEC-targeting nanoparticle-peptide-carriers abrogated CD4+ and CD8+ T cell immune responses towards the targeted peptide-epitopes and significantly improved transgene persistence in liver and muscle. Furthermore, nanoparticles carrying a single peptide-epitope recognized by anti-transgene CD8+ T cells also induced tolerance in CD8+ and CD4+ T cells specific for other transgene- and capsid-derived peptide-epitopes. This remarkable extension of tolerance, which combines linked suppression and infectious tolerance, is reported here for the first time and termed tolerance-spreading (Tspread). Importantly, Tspread remained restricted to the antigens carried by the AAV vectors and did not impair concurrent vaccination against an unrelated third-party tumour antigen.
Conclusion: Reported for the first time in immunotherapy, nanoparticle-peptide-carriers targeting LSECs induce Tspread to other relevant capsid- and transgene-derived epitopes, which is beneficial for achieving robust and durable tolerance in vivo. This finding is of great clinical importance, offering a novel antigen-specific strategy to improve viral vector-mediated gene transfer.