A Toll-like receptor recognizes bacterial DNA (original) (raw)

Nature volume 408, pages 740–745 (2000)Cite this article

An Erratum to this article was published on 01 February 2001

Abstract

DNA from bacteria has stimulatory effects on mammalian immune cells1,2,3, which depend on the presence of unmethylated CpG dinucleotides in the bacterial DNA. In contrast, mammalian DNA has a low frequency of CpG dinucleotides, and these are mostly methylated; therefore, mammalian DNA does not have immuno-stimulatory activity. CpG DNA induces a strong T-helper-1-like inflammatory response4,5,6,7. Accumulating evidence has revealed the therapeutic potential of CpG DNA as adjuvants for vaccination strategies for cancer, allergy and infectious diseases8,9,10. Despite its promising clinical use, the molecular mechanism by which CpG DNA activates immune cells remains unclear. Here we show that cellular response to CpG DNA is mediated by a Toll-like receptor, TLR9. TLR9-deficient (TLR9-/-) mice did not show any response to CpG DNA, including proliferation of splenocytes, inflammatory cytokine production from macrophages and maturation of dendritic cells. TLR9-/- mice showed resistance to the lethal effect of CpG DNA without any elevation of serum pro-inflammatory cytokine levels. The in vivo CpG-DNA-mediated T-helper type-1 response was also abolished in TLR9-/- mice. Thus, vertebrate immune systems appear to have evolved a specific Toll-like receptor that distinguishes bacterial DNA from self-DNA.

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References

  1. Krieg, A. M. Lymphocyte activation by CpG dinucleotide motifs in prokaryotic DNA. Trends Microbiol. 4, 73–76 (1996).
    Article CAS PubMed Google Scholar
  2. Lipford, G. B., Heeg, K. & Wagner, H. Bacterial DNA as immune cell activator. Trends Microbiol. 6, 496–500 (1998).
    Article CAS PubMed Google Scholar
  3. Yamamoto, S., Yamamoto, T. & Tokunaga, T. The discovery of immunostimulatory DNA sequence. Spring. Ser. Immunopathol. 22, 11–19 (2000).
    Article CAS Google Scholar
  4. Jakob, T., Walker, P. S., Krieg, A. M., Udey, M. C. & Vogel, J. C. Activation of cutaneous dendritic cells by CpG-containing oligodeoxynucleotides: a role for dendritic cells in the augmentation of Th1 responses by immunostimulatory DNA. J. Immunol. 161, 3042–3049 (1998).
    CAS PubMed Google Scholar
  5. Sparwasser, T. et al. Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells. Eur. J. Immunol. 28, 2045–2054 (1998).
    Article CAS PubMed Google Scholar
  6. Hartmann, G., Weiner, G. J. & Krieg, A. M. CpG DNA: a potent signal for growth, activation, and maturation of human dendritic cells. Proc. Natl Acad. Sci. USA 96, 9305–9310 (1999).
    Article ADS CAS PubMed PubMed Central Google Scholar
  7. Häcker, H. et al. Cell type-specific activation of mitogen-activated protein kinases by CpG-DNA controls interleukin-12 release from antigen-presenting cells. EMBO J. 18, 6973–6982 (1999).
    Article PubMed PubMed Central Google Scholar
  8. Wagner, H. Bacterial CpG DNA activates immune cells to signal infectious danger. Adv. Immunol. 73, 329–368 (1999).
    Article CAS PubMed Google Scholar
  9. Klinman, D. M., Verthelyi, D., Takeshita, F. & Ishii, K. J. Immune recognition of foreign DNA: a cure for bioterrorism? Immunity 11, 123–129 (1999).
    Article CAS PubMed Google Scholar
  10. Krieg, A. M. The role of CpG motifs in innate immunity. Curr. Opin. Immunol. 12, 35–43 (2000).
    Article CAS PubMed Google Scholar
  11. Medzhitov, R. & Janeway, C. A. Jr Innate immunity: the virtues of a nonclonal system of recognition. Cell 91, 295–298 (1997).
    Article CAS PubMed Google Scholar
  12. Medzhitov, R., Preston-Hurlburt, P. & Janeway, C. A. Jr A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature 388, 394–397 (1997).
    Article ADS CAS PubMed Google Scholar
  13. Rock, F. L., Hardiman, G., Timans, J. C., Kastelein, R. A. & Bazan, J. F. A family of human receptors structurally related to Drosophila Toll. Proc. Natl Acad. Sci. USA 95, 588–593 (1998).
    Article ADS CAS PubMed PubMed Central Google Scholar
  14. Takeuchi, O. et al. TLR6: A novel member of an expanding toll-like receptor family. Gene 231, 59–65 (1999).
    Article CAS PubMed Google Scholar
  15. Poltorak, A. et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085–2088 (1998).
    Article ADS CAS PubMed Google Scholar
  16. Hoshino, K. et al. Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J. Immunol. 162, 3749–3752 (1999).
    CAS PubMed Google Scholar
  17. Yoshimura, A. et al. Recognition of Gram-positive bacterial cell wall components by the innate immune system occurs via Toll-like receptor 2. J. Immunol. 163, 1–5 (1999).
    CAS PubMed Google Scholar
  18. Brightbill, H. D. et al. Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors. Science 285, 732–736 (1999).
    Article CAS PubMed Google Scholar
  19. Aliprantis, A. O. et al. Cell activation and apoptosis by bacterial lipoproteins through Toll-like receptor-2. Science 285, 736–739 (1999).
    Article CAS PubMed Google Scholar
  20. Underhill, D. M. et al. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature 401, 811–815 (1999).
    Article ADS CAS PubMed Google Scholar
  21. Takeuchi, O. et al. Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive bacterial cell wall components. Immunity 11, 443–451 (1999).
    Article CAS PubMed Google Scholar
  22. Takeuchi, O. et al. Preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2- and MyD88-dependent signaling pathway. J. Immunol. 164, 554–557 (2000).
    Article CAS PubMed Google Scholar
  23. Kawai, T., Adachi, O., Ogawa, T., Takeda, K. & Akira, S. Unresponsiveness of MyD88-deficient mice to endotoxin. Immunity 11, 115–122 (1999).
    Article CAS PubMed Google Scholar
  24. Häcker, H. et al. Immune cell activation by bacterial CpG-DNA through myeloid differential marker 88 and tumor necrosis factor receptor-associated factor (TRAF)6. J. Exp. Med. 192, 595–600 (2000).
    Article PubMed PubMed Central Google Scholar
  25. Sparwasser, T. et al. Macrophages sense pathogens via DNA motifs: induction of tumor necrosis factor-α-mediated shock. Eur. J. Immunol. 27, 1671–1679 (1997).
    Article CAS PubMed Google Scholar
  26. Lipford, G. B. et al. CpG-DNA-mediated transient lymphadenopathy is associated with a state of Th1 predisposition to antigen-driven responses. J. Immunol. 165, 1228–1235 (2000).
    Article CAS PubMed Google Scholar
  27. Liang, H., Reich, C. F., Pisetsky, D. S., Lipsky, P. E. The role of cell surface receptors in the activation of human B cells by phosphorothioate oligonucleotides. J. Immunol. 165, 1438–1445 (2000).
    Article CAS PubMed Google Scholar
  28. Krieg, A. M. et al. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 374, 546–549 (1995).
    Article ADS CAS PubMed Google Scholar
  29. Macfarlane, D. E. & Manzel, L. Antagonism of immunostimulatory CpG-oligodeoxynucleotides by quinacrine, chloroquine, and structurally related compounds. J. Immunol. 160, 1122–1131 (1998).
    CAS PubMed Google Scholar
  30. Häcker, H. et al. CpG-DNA-specific activation of antigen-presenting cells requires stress kinase activity and is preceded by non-specific endocytosis and endosomal maturation. EMBO J. 17, 6230–6240 (1998).
    Article PubMed PubMed Central Google Scholar
  31. Yi, A. K. & Krieg, A. M. Rapid induction of mitogen-activated protein kinases by immune stimulatory CpG DNA. J. Immunol. 161, 4493–4497 (1998).
    CAS PubMed Google Scholar

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Acknowledgements

We thank G. B. Lipford for helpful discussions; T. Aoki for secretarial assistance; and N. Tsuji, N. Iwami and E. Nakatani for technical assistance. We also thank Hayashibara Biochemical Laboratories, Inc. for providing anti-IRAK antibody. This work was supported in part by grants from the Ministry of Education, Science, Sports and Culture of Japan, and Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.

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Authors and Affiliations

  1. Department of Host Defense, Research Institute for Microbial Diseases, Osaka University,
    Hiroaki Hemmi, Osamu Takeuchi, Taro Kawai, Tsuneyasu Kaisho, Shintaro Sato, Hideki Sanjo, Makoto Matsumoto, Katsuaki Hoshino, Kiyoshi Takeda & Shizuo Akira
  2. Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 3-1 Yamada-oka, Suita, 565-0871, Osaka, Japan
    Hiroaki Hemmi, Osamu Takeuchi, Taro Kawai, Tsuneyasu Kaisho, Shintaro Sato, Hideki Sanjo, Makoto Matsumoto, Katsuaki Hoshino, Kiyoshi Takeda & Shizuo Akira
  3. Institute of Medical Microbiology, Immunology and Hygiene, Technical University of Munich, Trogerstr. 9, Munich, D-81675, Germany
    Hermann Wagner

Authors

  1. Hiroaki Hemmi
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  2. Osamu Takeuchi
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  3. Taro Kawai
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  4. Tsuneyasu Kaisho
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  5. Shintaro Sato
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  9. Hermann Wagner
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  10. Kiyoshi Takeda
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  11. Shizuo Akira
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Correspondence toShizuo Akira.

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Hemmi, H., Takeuchi, O., Kawai, T. et al. A Toll-like receptor recognizes bacterial DNA.Nature 408, 740–745 (2000). https://doi.org/10.1038/35047123

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