Preclinical evaluation of a recombinant adeno-associated virus vector expressing human alpha-1 antitrypsin made using a recombinant herpes simplex virus production method

UMMS Affiliation

Department of Pediatrics; Gene Therapy Center

Publication Date


Document Type



Analysis of Variance; Animals; Cells, Cultured; Dependovirus; Drug Evaluation, Preclinical; Female; Gene Therapy; Genetic Vectors; HEK293 Cells; Humans; Injections, Intramuscular; Male; Mice; Mice, Inbred C57BL; Muscle, Skeletal; Plasmids; Simplexvirus; Transfection; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency


Allergy and Immunology | Genetics and Genomics | Pediatrics


Recombinant adeno-associated virus (rAAV) vectors offer promise for gene therapy of alpha-1 antitrypsin (AAT) deficiency. A toxicology study in mice evaluated intramuscular injection of an rAAV vector expressing human AAT (rAAV-CB-hAAT) produced using a herpes simplex virus (HSV) complementation system or a plasmid transfection (TFX) method at doses of 3 x 10(11) vg (1.2 x 10(13) vg/kg) for both vectors and 2 x 10(12) vg (8 x 10(13) vg/kg) for the HSV-produced vector. The HSV-produced vector had favorable in vitro characteristics in terms of purity, efficiency of transduction, and hAAT expression. There were no significant differences in clinical findings or hematology and clinical chemistry values between test article and control groups and no gross pathology findings. Histopathological examination demonstrated minimal to mild changes in skeletal muscle at the injection site, consisting of focal chronic interstitial inflammation and muscle degeneration, regeneration, and vacuolization, in vector-injected animals. At the 3 x 10(11) vg dose, serum hAAT levels were higher with the HSV-produced vector than with the TFX-produced vector. With the higher dose of HSV-produced vector, the increase in serum hAAT levels was dose-proportional in females and greater than dose-proportional in males. Vector copy numbers in blood were highest 24 hr after dosing and declined thereafter, with no detectable copies present 90 days after dosing. Antibodies to hAAT were detected in almost all vector-treated animals, and antibodies to HSV were detected in most animals that received the highest vector dose. These results support continued development of rAAV-CB-hAAT for treatment of AAT deficiency.

DOI of Published Version



Hum Gene Ther. 2011 Feb;22(2):155-65. Epub 2010 Dec 12. Link to article on publisher's site

Journal/Book/Conference Title

Human gene therapy

Related Resources

Link to Article in PubMed

PubMed ID