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DNA Vaccines pp 173–184Cite as

In Vitro Assay of Immunostimulatory Activities of Plasmid Vectors

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Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 29))

Abstract

DNA vaccination represents a powerful new approach for the elicitation of long-lived protective immunity against a broad range of protein antigens (1,2). In this approach, the vaccine is a plasmid DNA vector that encodes a foreign protein to be targeted for the induction of humoral or cellular responses. Following administration by various routes, the plasmid is taken up by cells to allow intracellular production of the protein for presentation to the immune system. Although the trafficking of the plasmid and its protein product is not well understood, the generation of responses ultimately involves a bone marrow-derived antigen presenting cell (3).

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References

  1. Vogel, F. R. and Sarver, N. (1995) Nucleic acid vaccines. Clin. Microbiol. Rev. 8, 406–410.

    PubMed  CAS  Google Scholar 

  2. Pardoll, D. M. and Beckerleg, A. M. (1995) Exposing the immunology of naked DNA vaccines. Immunity 3, 165–169.

    Article  PubMed  CAS  Google Scholar 

  3. Corr, M., Lee, D. J., Carson, D. A., and Tighe, H. (1996) Gene vaccination with naked plasmid DNA: mechanism of CTL priming. J. Exp. Med. 184, 1555–1560.

    Article  PubMed  CAS  Google Scholar 

  4. Pisetsky, D. S. (1996) The immunologic properties of DNA. J. Immunol. 156, 421–423.

    PubMed  CAS  Google Scholar 

  5. Pisetsky, D. S. (1996) Immune activation by bacterial DNA: a new genetic code. Immunity 5, 303–310.

    Article  PubMed  CAS  Google Scholar 

  6. Yamamoto, S., Kuramoto, E., Shimada, S., and Tokunaga, T. (1988) In vitro augmentation of natural killer cell activity and production of interferon-α/ß and-γ with deoxyribonucleic acid fraction from Mycobacterium bovis BCG. Jpn. J. Cancer Res. 79, 866–873.

    PubMed  CAS  Google Scholar 

  7. Yamamoto, S., Yamamoto, T., Shimada, S., Kuramoto, E., Yano, O., Kataoka, T., and Tokunaga, T. (1992) DNA from bacteria, but not from vertebrates, induces interferons, activates natural killer cells and inhibits tumor growth. Microbiol. Immunol. 36, 983–997.

    PubMed  CAS  Google Scholar 

  8. Messina, J. P., Gilkeson, G. S., and Pisetsky, D. S. (1991) Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA. J. Immunol. 147, 1759–1764.

    PubMed  CAS  Google Scholar 

  9. Krieg, A. M., Yi, A.-K., Matson, S., Waldschmidt, T. J., Bishop, G. A., Teasdale, R., et al. (1995) CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 374, 546–549.

    Article  PubMed  CAS  Google Scholar 

  10. Klinman, D. M., Yi, A.-K., Beaucage, S. L., Conover, J., and Krieg, A. M. (1996) CpG motifs present in bacterial DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon γ. Proc. Natl. Acad. Sci. USA 93, 2879–2883.

    Article  PubMed  CAS  Google Scholar 

  11. Halpern, M. D., Kurlander, R. J., and Pisetsky, D. S. (1996) Bacterial DNA induces murine interferon-γ production by stimulation of interleukin-12 and tumor necrosis factor-α. Cell. Immunol. 167, 72–78.

    Article  PubMed  CAS  Google Scholar 

  12. Stacey, K. J., Sweet, M. J., and Hume, D. A. (1996) Macrophages ingest and are activated by bacterial DNA. J. Immunol. 157, 2116–2122.

    PubMed  CAS  Google Scholar 

  13. Yi, A.-K., Klinman, D. M., Martin, T. L., Matson, S., and Krieg, A. M. (1996) Rapid immune activation by CpG motifs in bacterial DNA. Systemic induction of IL-6 transcription through an antioxidant-sensitive pathway. J. Immunol. 157, 5394–5402.

    PubMed  CAS  Google Scholar 

  14. Kataoka, T., Yamamoto, S., Yamamoto, T., Kuramoto, E., Kimura, Y., Yano, O., and Tokunaga, T. (1992) Antitumor activity of synthetic oligonucleotides with sequences from cDNA encoding proteins of Mycobacterium bovis BCG. Jpn. J. Cancer Res. 83, 244–247.

    PubMed  CAS  Google Scholar 

  15. Krieg, A. M. (1995) CpG DNA: a pathogenic factor in systemic lupus erythematosus? J. Clin. Immunol. 15, 284–292.

    Article  PubMed  CAS  Google Scholar 

  16. Messina, J. P., Gilkeson, G. S., and Pisetsky, D. S. (1993) The influence of DNA structure on the in vitro stimulation of murine lymphocytes by natural and synthetic polynucleotide antigens. Cell. Immunol. 147, 148–157.

    Article  PubMed  CAS  Google Scholar 

  17. Pisetsky, D. S. and Reich, C. (1993) Stimulation of in vitro proliferation of murine lymphocytes by synthetic oligodeoxynucleotides. Molec. Biol. Rep. 18, 217–221.

    Article  CAS  Google Scholar 

  18. Kimura, Y., Sonehara, K., Kuramoto, E., Makino, T., Yamamoto, S., Yamamoto, T., et al. (1994) Binding of oligoguanylate to scavenger receptors is required for oligonucleotides to augment NK cell activity and induce IFN. J. Biochem. 116, 991–994.

    PubMed  CAS  Google Scholar 

  19. Krieger, M. and Hers, J. (1994) Structures and functions of multiligand lipoprotein receptors: macrophage scavenger receptors and LDL receptor-related protein (LRP). Annu. Rev. Biochem. 63, 604–637.

    Article  Google Scholar 

  20. Sato, Y., Roman, M., Tighe, H., Lee, D., Corr, M., Nguyen, M.-D., Silverman, G. J., Lotz, M., Carson, D. A., and Raz, E. (1996) Immunostimulatory DNA sequences necessary for effective intradermal gene immunization. Science 273, 352–354.

    Article  PubMed  CAS  Google Scholar 

  21. Klinman, D. M., Yamshchikov, G., and Ishigatsubo, Y. (1997) Contribution of CpG motifs to the immunogenicity of DNA vaccines. J. Immunol. 158, 3635–3639.

    PubMed  CAS  Google Scholar 

  22. Raz, E., Tighe, H., Sato, Y., Corr, M., Dudler, J. A., Roman, M., et al. (1996) Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization. Proc. Natl. Acad. Sci. USA 93, 5141–5145.

    Article  PubMed  CAS  Google Scholar 

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© 2000 Humana Press Inc., Totowa, NJ

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Reich, C.F., Pisetsky, D.S. (2000). In Vitro Assay of Immunostimulatory Activities of Plasmid Vectors. In: Lowrie, D.B., Whalen, R.G. (eds) DNA Vaccines. Methods in Molecular Medicine™, vol 29. Humana Press. https://doi.org/10.1385/1-59259-688-6:173

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  • DOI: https://doi.org/10.1385/1-59259-688-6:173

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-580-5

  • Online ISBN: 978-1-59259-688-1

  • eBook Packages: Springer Protocols

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