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Alphavirus-based DNA vaccine breaks immunological tolerance by activating innate antiviral pathways

Nature Medicine volume 9, pages 33–39 (2003)Cite this article

Abstract

Cancer vaccines targeting 'self' antigens that are expressed at consistently high levels by tumor cells are potentially useful in immunotherapy, but immunological tolerance may block their function. Here, we describe a novel, naked DNA vaccine encoding an alphavirus replicon (self-replicating mRNA) and the self/tumor antigen tyrosinase-related protein-1. Unlike conventional DNA vaccines, this vaccine can break tolerance and provide immunity to melanoma. The vaccine mediates production of double-stranded RNA, as evidenced by the autophosphorylation of dsRNA-dependent protein kinase R (PKR). Double-stranded RNA is critical to vaccine function because both the immunogenicity and the anti-tumor activity of the vaccine are blocked in mice deficient for the RNase L enzyme, a key component of the 2′,5′-linked oligoadenylate synthetase antiviral pathway involved in double-stranded RNA recognition. This study shows for the first time that alphaviral replicon-encoding DNA vaccines activate innate immune pathways known to drive antiviral immune responses, and points the way to strategies for improving the efficacy of immunization with naked DNA.

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Figure 1: Design of plasmids used for this study.
Figure 2: Antigen expression does not correlate with immunogenicity.
Figure 3: Tumor prevention by immunization with various plasmids encoding TRP-1.
Figure 4: Production of biologically active dsRNA in cells transfected with replicase-based plasmids.
Figure 5: The involvement of dsRNA and dsRNA-dependent pathways in the immune response to replicase-based plasmids.
Figure 6: The involvement of dsRNA and dsRNA-dependent pathways in the immune response after immunization with replicase-based plasmids.

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Acknowledgements

We thank L. Finch for FACS sorting, and E.S. Bergmann-Leitner and S. Frank for critical review of the manuscript. Supported in part by grant R01-AI34039 (to B.R.G.W.) and CA44059 (to R.H.S.) from the National Institutes of Health.

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Authors and Affiliations

  1. National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Wolfgang W. Leitner, Leroy N. Hwang, Han Ying & Nicholas P. Restifo

  2. Department of Cancer Biology, Cleveland Clinic Foundation, Cleveland, Ohio, USA

    Michael J. deVeer, Robert H. Silverman & Bryan R.G. Williams

  3. Department of Chemistry, Clinical Chemistry Program, Cleveland State University, Cleveland, Ohio, USA

    Aimin Zhou

  4. Center for Gene Therapy, Chiron Corporation, Emeryville, California, USA

    Thomas W. Dubensky

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  1. Wolfgang W. Leitner
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Correspondence to Wolfgang W. Leitner or Nicholas P. Restifo.

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Leitner, W., Hwang, L., deVeer, M. et al. Alphavirus-based DNA vaccine breaks immunological tolerance by activating innate antiviral pathways. Nat Med 9, 33–39 (2003). https://doi.org/10.1038/nm813

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