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

被引:72
作者
Virella-Lowell, I
Poirier, A
Chesnut, KA
Brantly, M
Flotte, TR
机构
[1] Univ Florida, Coll Med, Powell Gene Therapy Ctr, Gainesville, FL 32610 USA
[2] Univ Florida, Coll Med, Dept Pediat, Gainesville, FL 32610 USA
[3] Univ Florida, Coll Med, Dept Med, Gainesville, FL 32610 USA
[4] Univ Florida, Coll Med, Dept Mol Genet & Microbiol, Gainesville, FL 32610 USA
关键词
gene therapy; adeno-associated virus; cystic fibrosis; alpha-1; antitrypsin; inflammation;
D O I
10.1038/sj.gt.3301268
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 adenoassociated 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-l-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 rAA V transduction, and that these effects may be reversible by AAT.
引用
收藏
页码:1783 / 1789
页数:7
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