Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model

Alice Tomei, Melanie M. Choe, Melody A. Swartz

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Asthmatic patients are more susceptible to viral infection, and we asked whether dynamic strain on the airway wall (such as that associated with bronchoconstriction) would influence the rate of viral infection of the epithelial and subepithelial cells. To address this, we characterized the barrier function of a three-dimensional culture model of the bronchial airway wall mucosa, modified the culture conditions for optimization of ciliogenesis, and compared epithelial and subepithelial green fluorescent protein (GFP) transduction by a pWpts-GFP lentivirus, pseudotyped with VSV-G, under static vs. dynamic conditions. The model consisted of human lung fibroblasts, bronchial epithelial cells, and a type I collagen matrix, and after 21 days of culture at air liquid interface, it exhibited a pseudostratified epithelium comprised of basal cells, mucus-secreting cells, and ciliated columnar cells with beating cilia. Microparticle tracking revealed partial coordination of mucociliary transport among groups of cells. Slow dynamic compression of the airway wall model (15% strain at 0.1 Hz over 3 days) substantially enhanced GFP transduction of epithelial cells and underlying fibroblasts. Fibroblast-only controls showed a similar degree of transduction enhancement when undergoing dynamic strain, suggesting enhanced transport through the matrix. Tight junction loss in the epithelium after mechanical stress was observed by immunostaining. We conclude that dynamic compressive strain such as that associated with bronchoconstriction may promote transepithelial transport and enhance viral transgene delivery to epithelial and subepithelial cells. This finding has significance for asthma pathophysiology as well as for designing delivery strategies of viral gene therapies to the airways.

Original languageEnglish
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume294
Issue number1
DOIs
StatePublished - Jan 1 2008
Externally publishedYes

Fingerprint

Epithelial Cells
Bronchoconstriction
Fibroblasts
Virus Diseases
Green Fluorescent Proteins
Epithelium
Mucociliary Clearance
Lentivirus
Mechanical Stress
Viral Genes
Cilia
Tight Junctions
Mucus
Collagen Type I
Transgenes
Genetic Therapy
Mucous Membrane
Asthma
Air
Lung

Keywords

  • Asthma
  • Bronchoconstriction
  • Ciliogenesis
  • Human
  • Mechanical stress
  • Three-dimensional model

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

Cite this

Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model. / Tomei, Alice; Choe, Melanie M.; Swartz, Melody A.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 294, No. 1, 01.01.2008.

Research output: Contribution to journalArticle

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