Partial correction of the CFTR-dependent ABPA mouse model with recombinant adeno-associated virus gene transfer of truncated CFTR gene

J Gene Med. 2008 Jan;10(1):51-60. doi: 10.1002/jgm.1119.

Abstract

Recently, we have developed a model of airway inflammation in a CFTR knockout mouse utilizing Aspergillus fumigatus crude protein extract (Af-cpe) to mimic allergic bronchopulmonary aspergillosis (ABPA) 1, an unusual IgE-mediated hypersensitivity syndrome seen in up to 15% of cystic fibrosis (CF) patients and rarely elsewhere. We hypothesized that replacement of CFTR via targeted gene delivery to airway epithelium would correct aberrant epithelial cytokine signaling and ameliorate the ABPA phenotype in CFTR-deficient (CFTR 489X - /-, FABP-hCFTR + / +) mice. CFTR knockout mice underwent intra-tracheal (IT) delivery of recombinant adeno-associated virus serotype 5 (rAAV5Delta-264CFTR) or rAAV5-GFP at 2.58 x 10(12) viral genomes/mouse. All mice were then sensitized with two serial injections (200 microg) of crude Af antigen via the intra-peritoneal (IP) route. Untreated mice were sensitized without virus exposure. Challenges were performed 2 weeks after final sensitization, using a 0.25% solution containing Aspergillus fumigatus crude protein extract delivered by inhalation on three consecutive days. The rAAV5Delta-264CFTR-treated mice had lower total serum IgE levels (172513 ng/ml +/- 1312) than rAAV5-GFP controls (26 892 ng/ml +/- 3715) (p = 0.037) and non-treated, sensitized controls (24 816 +/- 4219 ng/ml). Serum IgG1 levels also were lower in mice receiving the CFTR vector. Interestingly, splenocytes from rAAV5Delta-264CFTR-treated mice secreted less IL-13, INFg, TNFa, RANTES and GM-CSF after ConA stimulation. Gene therapy with rAAV5Delta-264CFTR attenuated the hyper-IgE response in this reproducible CF mouse model of ABPA, with systemic effects also evident in the cytokine response of stimulated splenocytes.

MeSH terms

  • Animals
  • Aspergillosis, Allergic Bronchopulmonary / chemically induced
  • Aspergillosis, Allergic Bronchopulmonary / genetics
  • Aspergillosis, Allergic Bronchopulmonary / therapy*
  • Aspergillus fumigatus
  • Cell Proliferation / drug effects
  • Complex Mixtures
  • Concanavalin A / pharmacology
  • Cystic Fibrosis Transmembrane Conductance Regulator / deficiency
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • Cystic Fibrosis Transmembrane Conductance Regulator / therapeutic use*
  • Cytokines / metabolism
  • Dependovirus / metabolism*
  • Disease Models, Animal
  • Fluorescein-5-isothiocyanate / metabolism
  • Gene Expression Regulation / drug effects
  • Genetic Therapy*
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunoglobulin E / blood
  • Immunologic Factors / metabolism
  • Mice
  • Mice, Inbred CFTR
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Mutation / genetics*
  • Spleen / cytology
  • Spleen / drug effects
  • Transduction, Genetic*
  • Transgenes

Substances

  • Complex Mixtures
  • Cytokines
  • Immunologic Factors
  • Mutant Proteins
  • Concanavalin A
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Green Fluorescent Proteins
  • Immunoglobulin E
  • Fluorescein-5-isothiocyanate