Premium
Partial correction of the CFTR‐dependent ABPA mouse model with recombinant adeno‐associated virus gene transfer of truncated CFTR gene
Author(s) -
Mueller Christian,
Torrez Daniel,
Braag Sofia,
Martino Ashley,
Clarke Tracy,
CampbellThompson Martha,
Flotte Terence R.
Publication year - 2008
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1119
Subject(s) - ovalbumin , cystic fibrosis , allergic bronchopulmonary aspergillosis , adeno associated virus , immunology , cytokine , recombinant dna , genetic enhancement , immunoglobulin e , microbiology and biotechnology , biology , immune system , medicine , gene , antibody , biochemistry , vector (molecular biology)
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 (rAAV5Δ‐264CFTR) or rAAV5‐GFP at 2.58 × 10 12 viral genomes/mouse. All mice were then sensitized with two serial injections (200 µg) 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 rAAV5Δ‐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 rAAV5Δ‐264CFTR‐treated mice secreted less IL‐13, INFg, TNFa, RANTES and GM‐CSF after ConA stimulation. Gene therapy with rAAV5Δ‐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. Copyright © 2007 John Wiley & Sons, Ltd.