Journal article
Repeat Dosing of AAV2.5T to Ferret Lungs Elicits an Antibody Response That Diminishes Transduction in an Age-Dependent Manner
Molecular therapy. Methods & clinical development, Vol.19, pp.186-200
12/11/2020
DOI: 10.1016/j.omtm.2020.09.008
PMCID: PMC7648090
PMID: 33209961
Abstract
Readministration of recombinant adeno-associated virus (rAAV) may be necessary to treat cystic fibrosis (CF) lung disease using gene therapy. However, little is known about rAAV-mediated immune responses in the lung. 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. AAV2.5T-SP183-hCFTRΔR efficiently transduced both human and ferret airway epithelial cultures and complemented CFTR Cl– currents in CF airway cultures. Delivery of AAV2.5T-hCFTRΔR to neonatal and juvenile ferret lungs produced hCFTR mRNA at 200%–300% greater levels than endogenous fCFTR. 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. 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. Notably, both age groups demonstrated a reduction in BALF anti-capsid binding immunoglobulin (Ig) G, IgM, and IgA antibodies after repeat dosing. Unique to juvenile ferrets was a suppression of plasma anti-capsid-binding IgM after the second vector administration. 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.
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The AAV2.5T vector evolved to transduce human-airway epithelium, but a preclinical animal model to evaluate this virus had not yet been identified. We show that AAV2.5T also effectively transduces ferret airway epithelium in vitro and lungs in vivo, but neutralizing antibodies limited the efficacy of repeat dosing in juvenile, but not neonatal, ferrets.
Details
- Title: Subtitle
- Repeat Dosing of AAV2.5T to Ferret Lungs Elicits an Antibody Response That Diminishes Transduction in an Age-Dependent Manner
- Creators
- Yinghua Tang - University of IowaZiying Yan - University of IowaShen Lin - Spirovant Science Inc., Philadelphia, PA 19104, USAEric D. Huntemann - Roy J. and Lucille A. Carver College of MedicineZehua Feng - University of IowaSoo-Yeun Park - University of IowaXingshen Sun - University of IowaEric Yuen - Spirovant Science Inc., Philadelphia, PA 19104, USAJohn F. Engelhardt - Roy J. and Lucille A. Carver College of Medicine
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Methods & clinical development, Vol.19, pp.186-200
- DOI
- 10.1016/j.omtm.2020.09.008
- PMID
- 33209961
- PMCID
- PMC7648090
- NLM abbreviation
- Mol Ther Methods Clin Dev
- ISSN
- 2329-0501
- eISSN
- 2329-0501
- Publisher
- Elsevier Inc
- Grant note
- DOI: 10.13039/100000002, name: National Institutes of Health, award: HL051670, DK047967, HL123482; name: Spirovant Sciences
- Language
- English
- Date published
- 12/11/2020
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984284332702771
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