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Journal of Virology, July 2009, p. 6817-6824, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.00278-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cytotoxic-T-Lymphocyte-Mediated Elimination of Target Cells Transduced with Engineered Adeno-Associated Virus Type 2 Vector In Vivo{triangledown}

Chengwen Li,1 Matt Hirsch,1 Nina DiPrimio,1,2 Aravind Asokan,1,3 Kevin Goudy,4 Roland Tisch,4 and R. Jude Samulski1,2*

Gene Therapy Center,1 Department of Pharmacology,2 Department of Genetics,3 Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 275994

Received 8 February 2009/ Accepted 6 April 2009

A recent clinical trial in patients with hemophilia B has suggested that adeno-associated virus (AAV) capsid-specific cytotoxic T lymphocytes (CTLs) eliminated AAV-transduced hepatocytes and resulted in therapeutic failure. AAV capsids elicit a CTL response in animal models; however, these capsid-specific CTLs fail to kill AAV-transduced target cells in mice. To better model the human clinical trial data in mice, we introduced an immunodominant epitope derived from ovalbumin (OVA; SIINFEKL) into the AAV capsid and tested CTL-mediated killing of AAV2-transduced target tissues in vivo. Initially, in vitro experiments demonstrated both classical class I and cross-presentation of the OVA antigen, following endogenous expression or AAV2-OVA vector transduction, respectively. Furthermore, an OVA-specific CTL response was elicited after muscular or systemic injection of the AAV2-OVA vector. Finally, CTL reactivity was enhanced in mice with established SIINFEKL-specific immunity after AAV2-OVA/{alpha}1 anti-trypsin (AAT) administration. Most importantly, these OVA-specific CTLs decreased AAT expression in mice treated with AAV2-OVA/AAT vector that followed a time course mimicking uncoating kinetics of AAV2 transduction in OVA-immunized mice. These results demonstrate that AAV capsid-derived antigens elicit CD8+ CTL reactivity, and these CTLs eliminated AAV-transduced target cells in mice. Notably, this model system can be exploited to study the kinetics of capsid presentation from different serotypes of AAV and permit the design of novel strategies to block CTL-mediated killing of AAV-transduced cells.

* Corresponding author. Mailing address: Gene Therapy Center, 7119 Thurston-Bowles, CB 7352, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599. Phone: (919) 962-3285. Fax: (919) 966-0907. E-mail: rjs{at}med.unc.edu

{triangledown} Published ahead of print on 15 April 2009.

Journal of Virology, July 2009, p. 6817-6824, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.00278-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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