Dual reporter comparative indexing of rAAV pseudotyped vectors in chimpanzee airway
Gene Therapy Center; Department of Pediatrics
Medical Subject Headings
Animals; Biological Evolution; Dependovirus; Gene Transfer Techniques; *Genes, Reporter; Genetic Vectors; Humans; Immune System; Luciferases; Pan troglodytes; Phylogeny; Respiratory System; Species Specificity; T-Lymphocytes; Transgenes
Allergy and Immunology | Genetics and Genomics | Pediatrics | Respiratory Tract Diseases
Selecting the most efficient recombinant adeno-associated virus (rAAV) serotype for airway gene therapy has been difficult due to cross-specific differences in tropism and immune response between humans and animal models. Chimpanzees--the closest surviving genetic relative of humans--provide a valuable opportunity to select the most effective serotypes for clinical trials in humans. However, designing informative experiments using this protected species is challenging due to limited availability and experimental regulations. We have developed a method using Renilla luciferase (RL) and firefly luciferase (FL) reporters to directly index the relative transduction and immune response of two promising rAAV serotypes following lung coinfection. Analysis of differential luciferase activity in chimpanzee airway brushings demonstrated a 20-fold higher efficiency for rAAV1 over rAAV5 at 90 days, a finding that was similar in polarized human airway epithelia. T-cell responses to AAV5 capsid were stronger than AAV1 capsid. This dual vector indexing approach may be useful in selecting lead vector serotypes for clinical gene therapy and suggests rAAV1 is preferred for cystic fibrosis.
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Citation: Mol Ther. 2010 Mar;18(3):594-600. Epub 2009 Oct 13. Link to article on publisher's site