Skip to main content
SpringerLink
Log in

The influence of base sequence on the immunostimulatory properties of DNA

  • Published:
Immunologic Research Aims and scope Submit manuscript

Abstract

DNA is a complex macromolecule whose immunological properties vary with base sequences. As shown with synthetic oligonucleotides, potent immune stimulation results from six base motifs called CpG motifs or immunostimulatory sequences (ISS). These sequences center on an unmethylated CpG dinucleotide and occur much more commonly in bacterial DNA than mammalian DNA. As such, CpG motifs may function as a danger signal to stimulate B cell activation and cytokine production. In addition to CpG motifs, runs of deoxyguanosine (dG) residues in DNA can induce B cell activation and promote macrophage cytokine expression by adjacent CpG motifs. The array of these sequences may determine the overall immune activity of a DNA molecule and affect such processes as host defense against infection as well as the use of plasmids and synthetic oligonucleotides to treat disease.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Tokunaga T, Yano 0, Kuramoto E, Kimura Y, Yamamoto T, Kataoka T, et al.: Synthetic oligonucleotides with particular base sequences from the cDNA encoding proteins ofMycobacterium bovis BCG induce interferons and activate natural killer cells. Microbiol Immunol 1992;36:55–66.

    PubMed  CAS  Google Scholar 

  2. Krieg AM, Yyi A-K, Matson S, Waldschmidt TJ, Bishop GA, Teasdale R, et al.: CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 1995; 374:546–549.

    Article  PubMed  CAS  Google Scholar 

  3. Pisetsky DS: Immunostimulatory DNA: a clear and present danger? Nature Med 1997;3:829–831.

    Article  PubMed  CAS  Google Scholar 

  4. Pisetsky DS: Immune activation by bacterial DNA: a new genetic code. Immunity 1996;5:303–310.

    Article  PubMed  CAS  Google Scholar 

  5. Pisetsky DS: The immunologic properties of DNA. J Immunol 1996;156:421–423.

    PubMed  CAS  Google Scholar 

  6. Tokunaga T, Yamamoto H, Shimada S, Abe H, Fukuda T, Fujisawa Y, et al.: Antitumor activity of deoxyribonucleic acid fraction fromMycobacterium bovis BCG. I. Isolation, physicochemical characterization, and antitumor activity. J Natl Can Inst 1984;72: 955–962.

    CAS  Google Scholar 

  7. Shimada S, Yano O, Inoue H, Kuramoto E, Fukuda T, Yamamoto H, et al.: Antitumor activity of the DNA fraction fromMycobacterium bovis BCG. II. Effects on various syngeneic mouse tumors. J Natl Can Inst 1985;74:681–688.

    CAS  Google Scholar 

  8. Braun W, Nakano M: Influence of oligodeoxyribonucleotides on early events in antibody formation. Proc Soc Exp Biol Med 1965; 119:701–707.

    PubMed  CAS  Google Scholar 

  9. Vogt W, Ruhl H, Wagner B, Diamantstein T: Stimulation of DNA synthesis in mouse lymphoid cells by polyanions in vitro. II. Relationship between adjuvant activity and stimulation of DNA synthesis by polyanions. Eur J Immunol 1973;3:493–496.

    Article  PubMed  CAS  Google Scholar 

  10. Yamamoto S, Yamamoto T, Kataoka T, Kuramoto E, Yano O, Tokunaga T: Unique palindromic sequences in synthetic oligonucleotides are required to induce INF and augment INF-mediated natural killer activity. J Immunol 1992; 148:4072–4076.

    PubMed  CAS  Google Scholar 

  11. Yamamoto T, Yamamoto S, Kataoka T, Komuro K, Kohase M, Tokunaga T: Synthetic oligonucleotides with certain palindromes stimulate inteferon production of human peripheral blood lymphocytesin vitro. Jpn J Cancer Res 1994;85:775–779.

    PubMed  CAS  Google Scholar 

  12. Yamamoto S, Yamamoto T, Shimada S, Kuramoto E, Yano O, Kataoka T, et al.: DNA from bacteria, but not from vertebrates, induces interferons, activates natural killer cells and inhibits tumor growth. Microbiol Immunol 1992;36:983–997.

    PubMed  CAS  Google Scholar 

  13. Messina JP, Gilkeson GS, Pisetsky DS: Stimulation ofin vitro murine lymphocyte proliferation by bacterial DNA. J Immunol 1991; 147:1759–1764.

    PubMed  CAS  Google Scholar 

  14. Halpern MD, Kurlander RJ, Pisetsky DS: Bacterial DNA induces murine interferon-γ production by stimulation of interleukin-12 and tumor necrosis factor-α. Cell Immunol 1996;167:72–78.

    Article  PubMed  CAS  Google Scholar 

  15. Klinman DM, Yi A-K, Beaucage SL, Conover J, Krieg AM: CpG motifs presents in bacterial DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon γ. Proc Natl Acad Sci USA 1996;93:2879–2883.

    Article  PubMed  CAS  Google Scholar 

  16. Cowdery JS, Chace JH, Yi A-K, Krieg AM: Bacterial DNA induces NK cells to produce IFN-γ in vivo and increases the toxicity of lipopolysaccharides. J Immunol 1996; 156:4570–4575.

    PubMed  CAS  Google Scholar 

  17. Yi A-K, Chace JH, Cowdery JS, Krieg AM: IFN-γ promotes IL-6 and IgM secretion in response to CpG motifs in bacterial DNA and oligodeoxynucleotides. J Immunol 1996; 156:558–564.

    PubMed  CAS  Google Scholar 

  18. Liang H, Nishioka Y, Reich CF, Pisetsky DS, Lipsky PE: Activation of human B cells by phosphorothioate oligodeoxynucleotides. J Clin Invest 1996;98: 1119–1129.

    Article  PubMed  CAS  Google Scholar 

  19. Bird AP: CpG islands as gene markers in the vertebrate nucleus. Trends Genet 1987;3:342–346.

    Article  CAS  Google Scholar 

  20. Hergersberg M: Biological aspects of cytosine methylation in eukaryotic cells. Experientia 1991;47: 1171–1185.

    Article  PubMed  CAS  Google Scholar 

  21. Burge C, Campbell AM, Karlin S: Over- and under-representation of short oligonucleotides in DNA sequences. Proc Natl Acad Sci USA 1992;89:1358–1362.

    Article  PubMed  CAS  Google Scholar 

  22. Bird AP: Functions for DNA methylation in vertebrates. Cold Harbor Symp Quant Biol 1993; 58:281–285.

    CAS  Google Scholar 

  23. Doerfler W:Patterns ofDNA methylation—evolutionary vestiges of foreign DNA inacti vation as a host defense mechanism. A proposal. Biol Chem Hoppe-Seyler 1991; 372:557–564.

    PubMed  CAS  Google Scholar 

  24. Sun S, Beard C, Jaenisch R, Jones P, Sprent J: Mitogenicity of DNA from different organisms for murine B cells. J Immunol 1997; 159:3119–3125.

    PubMed  CAS  Google Scholar 

  25. Pisetsky DS, Reich C:Stimulation ofin vitro proliferation of murine lymphocytes by synthetic oligodeoxynucleotides. Mol Biol Rep 1993; 18:217–221.

    Article  PubMed  CAS  Google Scholar 

  26. Messina JP, Gilkeson GS, Pisetsky DS: The influence of DNA structure on thein vitro stimulation of murine lymphocytes by natural and synthetic polynucleotide antigens. Cell Immunol 1993;147:148–157.

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  28. Williamson JR: G-quartet structures in telomeric DNA. Annu Rev Biophys Biomol Struct 1994;23: 703–730.

    Article  PubMed  CAS  Google Scholar 

  29. Zakian VA: Telomeres: Beginning to understand the end. Science 1995;270:1601–1607.

    Article  PubMed  CAS  Google Scholar 

  30. Kodama T, Freeman M, Rohrer L, Zabrecky J, Matsudaira P, Krieger M: Type I macrophage scavenger receptor contains α-helical and collagen-like coiled coils. Nature 1990;343:531–535.

    Article  PubMed  CAS  Google Scholar 

  31. Krieger M, Herz J: Structures and functions of multiligand lipoprotein receptors: macrophage scavenger receptors andL DL receptor-related protein (LRP). Annu Rev Biochem 1994;63:601–637.

    PubMed  CAS  Google Scholar 

  32. Pearson AM, Rich A, Krieger M: Polynucleotide binding to macrophage scavenger receptors depends on the formation of basequartet-stabilized four-stranded helices. J Biol Chem 1993;268: 3546–3554.

    PubMed  CAS  Google Scholar 

  33. Haworth R, Platt N, Keshav S, Hughes D, Darley E, Suzuki H, et al.: The macrophage scavenger receptor type A is expressed by activated macrophages and protects the host against lethal endotoxic shock. J Exp Med 1997;186: 1431–1439.

    Article  PubMed  CAS  Google Scholar 

  34. Haberland ME, Rasmussen RR, Fogelman AM: Receptor recognition of maleyl-albumin induces chemotaxis in human monocytes. J Clin Invest 1986;78:827–831.

    PubMed  CAS  Google Scholar 

  35. Frostegard J, Nilsson J, Haegerstrand A, Hamsten A, Wigzell H, Gidlund M: Oxidized low density lipoprotein induces differentiation and adhesion of human monocytes and the monocytic cell line U937. Proc Natl Acad Sci 1990; 87:904–908.

    Article  PubMed  CAS  Google Scholar 

  36. Pisetsky DS, Reich CF: The influence of base sequence on the immunological properties of defined oligonucleotides. Immunopharmacology 1998;40:199–208.

    Article  PubMed  CAS  Google Scholar 

  37. Hughes JA, Avrutskaya AV, Juliano RL: Influence of base composition on membrane binding and cellular uptake of 10-mer phosphorothioate oligonucleotides in Chinese hamster ovary (CHRC5) cells. Antisense Res Dev 1994;4: 211–215.

    PubMed  CAS  Google Scholar 

  38. Yi A-K, Tuetken R, Redford T, Waldschmidt M, Kirsch J, Krieg AM: CpG motifs in bacterial DNA activate leukocytes through the pH-dependent generation of reactive oxygen species. J Immunol 1998;160:4755–4761.

    PubMed  CAS  Google Scholar 

  39. Vogel FR, Sarver N: Nucleic acid vaccines. Clin Microbiol Rev 1995; 8:406–410.

    PubMed  CAS  Google Scholar 

  40. Pardoll DM, Beckerleg AM: Exposing the immunology of naked DNA vaccines. Immunity 1995;3:165–169.

    Article  PubMed  CAS  Google Scholar 

  41. Corr M, Lee DJ, Carson DA, Tighe H: Gene vaccination with naked plasmid DNA: mechanism of CTL priming. J Exp Med 1996; 184: 1555–1560.

    Article  PubMed  CAS  Google Scholar 

  42. Iwasaki A, Torres CAT, Ohashi PS, Robinson HL, Barber BH: The dominant role of bone marrowderived cells in CTL induction following plasmid DNA immunization at different sites. J Immunol 1997;159:11–14.

    PubMed  CAS  Google Scholar 

  43. Sato Y, Roman M, Tighe H, Lee D, Corr M, Nguyen M-D, et al.: Immunostimulatory DNA sequences necessary for effective intradermal gene immunization. Science 1996;273:352–354.

    Article  PubMed  CAS  Google Scholar 

  44. Klinman DM, Yamshchikov G, Ishigatsubo Y: Contribution of CpG motifs to the immunogenicity of DNA vaccines. J Immunol 1997;158:3635–3639.

    PubMed  CAS  Google Scholar 

  45. Wloch MK, Pasquini S, Ertl HCJ, Pisetsky DS: The influence of DNA sequence on the immunostimulatory properties of plasmid DNA vectors. Hum Gene Ther 1998;9:1439–1447.

    PubMed  CAS  Google Scholar 

  46. Kawabata K, Takakura Y, Hashida M: The fate of plasmid DNA after intravenous injection in mice: involvement of scavenger receptors in its hepatic uptake. Pharm Res 1995;12:825–830.

    Article  PubMed  CAS  Google Scholar 

  47. Sawai K, Mahato RI, Oka Y, Takakura Y, Hashida M: Diposition of oligonucleotides in isolated perfused rat kidney: involvement of scavenger receptors in their renal uptake. J Pharmacol Exp Ther 1996;279:284–290.

    PubMed  CAS  Google Scholar 

  48. Stein CA, Cheng YC: Antisense oligonucleotides as therapeutic agents—is the bullet really magical? Science 1993;261: 1004–1012.

    Article  PubMed  CAS  Google Scholar 

  49. Frey PA, Sammons RD: Bond order and charge localization in nucleosidephosphorothioates. Science 1985;228:541–545.

    Article  PubMed  CAS  Google Scholar 

  50. Stein CA, Subasinghe C, Shinozuka K, Cohen JS: Physicochemical properties of phosphorothioate oligodeoxynucleotides. Nucleic Acids Res 1988; 16:3209–3221.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  52. Lipford GB, Bauer M, Blank C, Reiter R, Wagner H, Heeg K: CpG-containing synthetic oligonucleotides promote B and cytotoxic T cell responses to protein antigen: a new class of vaccine adjuvants. Eur J Immunol 1997; 27:2340–2344.

    Article  PubMed  CAS  Google Scholar 

  53. Sun S, Kishimoto H, Sprent J: DNA as an adjuvant: capacity of insect DNA and synthetic oligodeoxynucleotides to augment T cell responses to specific antigen. J Exp Med 1998;187:1145–1150.

    Article  PubMed  CAS  Google Scholar 

  54. Zimmermann S, Egeter 0, Hausmann S, Lipford GB, Rocken M, Wagner H, et al.: Cutting Edge: CpG oligodeoxynucleotides trigger protective and curative Thl responses in lethal murine Leishmaniasis. J Immunol 1998; 160: 3627–3630.

    PubMed  CAS  Google Scholar 

  55. Agrawal S, Temsamani J, Galbraith W, Tang J: Pharmacokinetics of antisense oligonucleotides. Clin Pharmacokinet 1995;28:7–16.

    PubMed  CAS  Google Scholar 

  56. Zhao Q, Temsamani J, Iadarola PL, Jiang Z, Agrawal S: Effect of different chemically modified oligodeoxynucleotides on immune stimulation. Biochem Pharm 1996; 51:173–182.

    Article  PubMed  CAS  Google Scholar 

  57. Monteith DK, Henry SP, Howard RB, Flournoy S, Levin AA, Bennett CF, et al.: Immune stimulation —a class effect of phosphorothioate oligodeoxynucleotides in rodents. Anticancer Drug Design 1997; 12:421–432.

    CAS  Google Scholar 

  58. Yakubov L, Khaled Z, Zhang L-M, Truneh A, Vlassov V, Stein CA: Oligodeoxynucleotides interact with recombinant CD4 at multiple sites. J Biol Chem 1993;268: 18,818–18,823.

    CAS  Google Scholar 

  59. Brown DA, Kang S-H, Gryaznov SM, Dedionisio L, Heidenreich 0, Sullivan S, et al.: Effect of phosphorothioate modification of oligodeoxynucleotides on specific protein binding. J Biol Chem 1994; 269:26,801–26,805.

    CAS  Google Scholar 

  60. Benimetskaya L, Loike JD, Khaled Z, Loike G, Silverstein SC, Cao L, et al.: Mac-1 (CD11b/CD18) is an oligodeoxynucleotide-binding protein. Nature Med 1997;3:414–420.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medical Service Durham VA Medical Center and Division of Rheumatology, Allergy and Clinical Immunology, Duke University Medical Center, Box 15IG, 508 Fulton St., 27705, Durham, NC

    David S. Pisetsky

Authors
  1. David S. Pisetsky
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David S. Pisetsky.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pisetsky, D.S. The influence of base sequence on the immunostimulatory properties of DNA. Immunol Res 19, 35–46 (1999). https://doi.org/10.1007/BF02786475

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02786475

Key Words

Search

Navigation