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Inhibition of recombinant adeno-associated virus (rAAV) transduction by bronchial secretions from cystic fibrosis patients

Gene Therapy volume 7, pages 1783–1789 (2000)Cite this article

Abstract

The conducting airways are the primary target for gene transfer in cystic fibrosis (CF), yet the inflammation associated with CF lung disease could potentially pose a significant barrier to gene transfer vectors, such as recombinant adeno-associated virus (rAAV). In order to investigate this possibility, aliquots of bronchoalveolar lavage (BAL) fluid from eight individuals with CF were tested for their in vitro inhibitory effects on rAAV transduction, along with BAL from non-CF individuals. While the non-CF BAL fluid was not inhibitory, seven of eight CF BAL samples had significant inhibitory activity, resulting in a five- to 20-fold reduction in transduction events. Inhibition of rAAV transduction by CF BAL could be reversed by alpha-1-antitrypsin (AAT), but not by DNase. When neutrophil elastase and neutrophil alpha defensins (human neutrophil peptides, HNP) were measured in these samples, they were elevated by 500- and 10000-fold, respectively. The levels of HNP correlated inversely with the amount of rAAV transduction. Furthermore, rAAV transduction could be blocked by purified HNP in an AAT-reversible manner at HNP concentrations within the range measured in these fluids. We conclude that products of inflammation in CF BAL fluid are inhibitory to rAAV transduction, and that these effects may be reversible by AAT.

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Acknowledgements

This work was supported by grants from NIH (HL51811, RR0082, DK51809). The original study from which the discard BAL was obtained was sponsored by Targeted Genetics Corporation. Many thanks to Margaret Humphries for help with study coordination and to Thomas Ferkol and Barrie Carter for advice that helped to guide these studies.

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Authors and Affiliations

  1. Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA

    I Virella-Lowell, A Poirier, K A Chesnut & T R Flotte

  2. Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA

    I Virella-Lowell, A Poirier & T R Flotte

  3. Department of Medicine, University of Florida College of Medicine, Gainesville, FL, USA

    M Brantly

  4. Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA

    K A Chesnut, M Brantly & T R Flotte

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  1. I Virella-Lowell
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Virella-Lowell, I., Poirier, A., Chesnut, K. et al. Inhibition of recombinant adeno-associated virus (rAAV) transduction by bronchial secretions from cystic fibrosis patients. Gene Ther 7, 1783–1789 (2000). https://doi.org/10.1038/sj.gt.3301268

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