In this study, we sought to evaluate the suitability of the ferret as a model for testing rAAV2.5T and humoral responses that could limit the efficacy of repeated dosing. Graphical AbstractĪnd thus, the ferret may be a suitable species for preclinical studies with rAAV2.5T. Thus, age-dependent immune system maturation and isotype switching may affect the development of high-affinity lung NAbs after repeat dosing of AAV2.5T and may provide a path to blunt AAV-neutralizing responses in the lung. Unique to juvenile ferrets was a suppression of plasma anti-capsid-binding IgM after the second vector administration. Notably, both age groups demonstrated a reduction in BALF anti-capsid binding immunoglobulin (Ig) G, IgM, and IgA antibodies after repeat dosing. Repeat dosing significantly reduced transgene expression (11-fold) and increased bronchoalveolar lavage fluid (BALF) NAbs only in juvenile, but not neonatal, ferrets, despite near-equivalent plasma NAb responses in both age groups. Single-dose (AAV2.5T-SP183-gLuc) or repeat dosing (AAV2.5T-SP183-fCFTRΔR followed by AAV2.5T-SP183-gLuc) of AAV2.5T was performed in neonatal and juvenile ferrets. Delivery of AAV2.5T-hCFTRΔR to neonatal and juvenile ferret lungs produced h CFTR mRNA at 200%–300% greater levels than endogenous f CFTR. AAV2.5T-SP183-hCFTRΔR efficiently transduced both human and ferret airway epithelial cultures and complemented CFTR Cl – currents in CF airway cultures. Here, we demonstrate the suitability of the ferret for testing AAV2.5T-mediated CFTR delivery to the lung and characterization of neutralizing-antibody (NAb) responses. However, little is known about rAAV-mediated immune responses in the lung. Readministration of recombinant adeno-associated virus (rAAV) may be necessary to treat cystic fibrosis (CF) lung disease using gene therapy.
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