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Journal of Virology, November 2002, p. 11343-11349, Vol. 76, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.22.11343-11349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A Limited Number of Transducible Hepatocytes Restricts a Wide-Range Linear Vector Dose Response in Recombinant Adeno-Associated Virus-Mediated Liver Transduction

Hiroyuki Nakai,1 Clare E. Thomas,1 Theresa A. Storm,1 Sally Fuess,1 Sharon Powell,2 J. Fraser Wright,2 and Mark A. Kay1*

Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California 94305,1 Avigen, Inc., Alameda, California 945022

Received 23 May 2002/ Accepted 7 August 2002

Recombinant adeno-associated virus (rAAV) vectors are promising vehicles for achieving stable liver transduction in vivo. However, the mechanisms of liver transduction are not fully understood, and furthermore, the relationships between rAAV dose and levels of transgene expression, total number of hepatocytes transduced, and proportion of integrated vector genomes have not been well established. To begin to elucidate the liver transduction dose response with rAAV vectors, we injected mice with two different human factor IX or Escherichia coli lacZ-expressing AAV serotype 2-based vectors at doses ranging between 4.0 x 108 and 1.1 x 1013 vector genomes (vg)/mouse, in three- to sixfold increments. A 2-log-range linear dose-response curve of transgene expression was obtained from 3.7 x 109 to 3.0 x 1011 vg/mouse. Vector doses above 3.0 x 1011 vg/mouse resulted in disproportionately smaller increases in both the number of transduced hepatocytes and levels of transgene expression, followed by saturation at doses above 1.8 x 1012 vg/mouse. In contrast, a linear increase in the number of vector genomes per hepatocyte was observed up to 1.8 x 1012 vg/mouse concomitantly with enhanced vector genome concatemerization, while the proportion of integrated vector genomes was independent of the vector dose. Thus, the mechanisms that restrict a wide-range linear dose response at high doses likely involve decreased functionality of vector genomes and restriction of transduction to fewer than 10% of total hepatocytes. Such information may be useful to determine appropriate vector doses for in vivo administration and provides further insights into the mechanisms of rAAV transduction in the liver.

* Corresponding author. Mailing address: Departments of Pediatrics and Genetics, 300 Pasteur Dr., Rm. G305A, Stanford University, Stanford, CA 94305. Phone: (650) 498-6531. Fax: (650) 498-6540. E-mail: markay{at}stanford.edu.

Journal of Virology, November 2002, p. 11343-11349, Vol. 76, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.22.11343-11349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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