Feline immunodeficiency virus vectors persistently transduce nondividing airway epithelia and correct the cystic fibrosis defect
Guoshun Wang, … , Beverly L. Davidson, Paul B. McCray Jr.
Guoshun Wang, … , Beverly L. Davidson, Paul B. McCray Jr.
Published December 1, 1999
Citation Information: J Clin Invest. 1999;104(11):R55-R62. https://doi.org/10.1172/JCI8390.
View: Text | PDF
Article

Feline immunodeficiency virus vectors persistently transduce nondividing airway epithelia and correct the cystic fibrosis defect

Abstract

Several problems limit the application of gene transfer to correct the cystic fibrosis (CF) Cl– transport defect in airway epithelia. These include inefficient transduction with vectors applied to the apical surface, a low rate of division by airway epithelial cells, failure of transgene expression to persist, and immune responses to vectors or vector-encoded proteins. To address these issues, we used a feline immunodeficiency virus–based (FIV-based) vector. FIV vector formulated with a calcium chelator transduced fully differentiated, nondividing human airway epithelia when applied to the apical surface. FIV-based vector encoding the cystic fibrosis transmembrane conductance regulator cDNA corrected the Cl– transport defect in differentiated CF airway epithelia for the life of the culture (>3 months). When this approach was applied in vivo, FIV vector expressing β-galactosidase transduced 1–14% of adult rabbit airway epithelia. Transduced cells were present in the conducting airways, bronchioles, and alveoli. Importantly, gene expression persisted, and cells with progenitor capacity were targeted. FIV-based lentiviral vectors may be useful for the treatment of genetic lung diseases such as CF.

Authors

Guoshun Wang, Vladimir Slepushkin, Joseph Zabner, Shaf Keshavjee, Julie C. Johnston, Sybille L. Sauter, Doug J. Jolly, Thomas W. Dubensky Jr., Beverly L. Davidson, Paul B. McCray Jr.

×

Figure 1

Options: View larger image (or click on image) Download as PowerPoint
FIV vectors transduce nondividing airway epithelia in vitro. (a) Applica...
FIV vectors transduce nondividing airway epithelia in vitro. (a) Application of FIV-βgal (moi 10) to the apical surface of the epithelial sheet resulted in no gene transfer. Representative en face view of X-gal stained epithelia 3 days after vector application. (b) FIV-βgal vector applied to airway epithelia from the basolateral surface in vitro (moi ∼10) resulted in gene transfer. Representative en face view is shown. (c) Gene transfer from the apical surface with VSV-G FIV is enhanced by physical disruption of epithelia. The epithelial sheet was scratched with a pipette tip prior to apical vector application. En face view shows gene transfer (gray cells) only along the area of epithelial disruption. (d) FIV transduces aphidicolin growth-arrested cells (en face view). Vector was applied to apical surface in the presence of 6 mM EGTA in hypotonic buffer. Approximately 17% of epithelia were transgene positive (range 12–22%; n = 5 epithelia from 2 different preparations). For cells in control media without aphidicolin, approximately 20% of cells expressed the transgene (data not shown; range 8–30%; n = 5 epithelia from 3 different preparations). (e) Quantification of gene transfer results under conditions shown in d (mean ± SEM; n = 5 epithelia; 3 different preparations).