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Enhanced transduction of mouse bone marrow-derived dendritic cells by repetitive infection with self-complementary adeno-associated virus 6 combined with immunostimulatory ligands

Gene Therapy volume 13, pages 29–39 (2006)Cite this article

Abstract

The potential of adeno-associated virus (AAV)-based vectors in human gene therapy is being explored for several diseases. Although sustained transgene expression and low vector-associated cellular immunity are attractive features of recombinant (r) AAV, the wider application of rAAV vectors encapsidated in serotype 2 capsid is hampered by poor transduction efficiency in many target tissues. These include ex vivo-generated dendritic cells (DC), which have demonstrated promising immunotherapeutic activity. We report here that efficient transduction of mouse bone marrow-derived DC can be achieved with self-complementary (sc) rAAV encapsidated in serotype 6 capsid. Sequential exposure of DC precursor cultures to IL-4 and GM-CSF with sc rAAV6 encoding the human tumor antigen, carcinoembryonic antigen (CEA), for 7 days followed by activation with CpG oligodeoxynucleotides (ODN) and anti-mouse CD40 antibody resulted in highly efficient transduction of DC. DC surface markers as determined by flow cytometry analysis of sc rAAV6-transduced DC were comparable to nontransduced DC. Efficiency of vector transduction and transgene expression were confirmed by immunostaining and real-time PCR. Microarray analysis of RNA from CpG ODN and CD40 antibody stimulated sc AAV6-transduced DC revealed upregulation of transcription factors and cytokines involved in immune activation and downregulation of inhibitory factors, suggesting a possible role of transcriptional activation in the observed effect. The adoptive transfer into syngeneic mice of the ex vivo-transduced and activated DC resulted in the development of CEA-specific antibody and T-helper 1-associated immune responses. Immunized mice also developed antibody to AAV6 capsid protein, which did not crossreact with AAV2 capsid protein. These studies demonstrate the potential utility of sc rAAV serotype 6-based vectors in transduction of DC for genetic vaccination approaches.

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Acknowledgements

Financial support from the National Institutes of Health Grants R01CA90850, R01CA98817 and P50 CA89019 is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Medicine, The University of Alabama at Birmingham, Birmingham, AL, USA

    W A Aldrich, C B Jenkins, D R Shaw, T V Strong & P L Triozzi

  2. The Comprehensive Cancer Center, The University of Alabama at Birmingham, Birmingham, AL, USA

    W A Aldrich, C B Jenkins, D R Shaw, T V Strong, P L Triozzi & S Ponnazhagan

  3. Department of Pathology, The University of Alabama at Birmingham, Birmingham, AL, USA

    C Ren, A F White, S Kumar & S Ponnazhagan

  4. Avigen Inc., Alameda, CA, USA

    S-Z Zhou

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  1. W A Aldrich
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Correspondence to S Ponnazhagan.

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Aldrich, W., Ren, C., White, A. et al. Enhanced transduction of mouse bone marrow-derived dendritic cells by repetitive infection with self-complementary adeno-associated virus 6 combined with immunostimulatory ligands. Gene Ther 13, 29–39 (2006). https://doi.org/10.1038/sj.gt.3302601

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