Immune-mediated destruction of transfected muscle fibers after direct gene transfer with antigen-expressing plasmid DNA (original) (raw)

Gene Therapy volume 4, pages 181–188 (1997)Cite this article

Abstract

DNA-based immunization of mice by intramuscular injection of antigen-encoding plasmid DNA results in immune responses which may be sustained for extended periods of time without an antigen boost. For example, we have previously shown that a strong humoral response against hepatitis B virus surface antigen (HBsAg) will persist for up to 74 weeks following a single intramuscular administration of DNA. It has been proposed that the longevity of the response is due to sustained expression of antigen in transfected muscle cells. However, here we show by immunohistochemistry and electron microscopy that HBsAg-expressing muscle fibers are destroyed around 10 days after injection of DNA in mice. We have also evaluated destruction of the transfected muscle fibers indirectly, by measurement of luciferase activity in muscles at different times after injection of a luciferase reporter gene construct, alone or in combination with HBsAg-expressing DNA. Control muscles injected with luciferase-expressing DNA alone maintain expression of high levels of luciferase for at least 60 days. In contrast, muscles co-injected with DNAs expressing luciferase and a secreted form of HBsAg show high levels of luciferase activity at 5 days but >99% of this is lost by 20 days. Similar results are obtained with co-expression of luciferase and β-galactosidase, a non-secreted antigen. Loss of luciferase expression does not occur in muscles of mice with severe combined immunodeficiency, indicating that the myofiber destruction is immunologically mediated.

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Authors and Affiliations

  1. Loeb Research Institute, Ottawa Civic Hospital, 1053 Carling Avenue, Ottawa, K1Y 4E9, Canada
    HL Davis & CL Brazolot Millan
  2. Program in Physiotherapy and Department of Physiology, University of Ottawa, Canada
    HL Davis
  3. Department of Cell Biology and Physiology, University of Pittsburgh, PA, USA
    SC Watkins

Authors

  1. HL Davis
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  2. CL Brazolot Millan
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  3. SC Watkins
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Davis, H., Millan, C. & Watkins, S. Immune-mediated destruction of transfected muscle fibers after direct gene transfer with antigen-expressing plasmid DNA.Gene Ther 4, 181–188 (1997). https://doi.org/10.1038/sj.gt.3300380

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