Article | Published:

Detection of pathogenic intestinal bacteria by Toll-like receptor 5 on intestinal CD11c+ lamina propria cells

Nature Immunology volume 7, pages 868874 (2006) | Download Citation

Subjects

Abstract

Toll-like receptors (TLRs) recognize distinct microbial components and induce innate immune responses. TLR5 is triggered by bacterial flagellin. Here we generated Tlr5−/− 1mice and assessed TLR5 function in vivo. Unlike other TLRs, TLR5 was not expressed on conventional dendritic cells or macrophages. In contrast, TLR5 was expressed mainly on intestinal CD11c+ lamina propria cells (LPCs). CD11c+ LPCs detected pathogenic bacteria and secreted proinflammatory cytokines in a TLR5-dependent way. However, CD11c+ LPCs do not express TLR4 and did not secrete proinflammatory cytokines after exposure to a commensal bacterium. Notably, transport of pathogenic Salmonella typhimurium from the intestinal tract to mesenteric lymph nodes was impaired in Tlr5−/− mice. These data suggest that CD11c+ LPCs, via TLR5, detect and are used by pathogenic bacteria in the intestinal lumen.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Gene Expression Omnibus

References

  1. 1.

    , & Pathogen recognition and innate immunity. Cell 124, 783–801 (2006).

  2. 2.

    et al. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099–1103 (2001).

  3. 3.

    Genetics and biogenesis of bacterial flagella. Annu. Rev. Genet. 26, 131–158 (1992).

  4. 4.

    , , , & Cutting edge: bacterial flagellin activates basolaterally expressed TLR5 to induce epithelial proinflammatory gene expression. J. Immunol. 167, 1882–1885 (2001).

  5. 5.

    & Salmonella flagellin, a microbial target of the innate and adaptive immune system. Immunol. Lett. 101, 117–122 (2005).

  6. 6.

    et al. Flagellin stimulation of intestinal epithelial cells triggers CCL20-mediated migration of dendritic cells. Proc. Natl. Acad. Sci. USA 98, 13722–13727 (2001).

  7. 7.

    et al. Cloning and characterization of the murine toll-like receptor 5 (Tlr5) gene: sequence and mRNA expression studies in Salmonella-susceptible MOLF/Ei mice. Genomics 64, 230–240 (2000).

  8. 8.

    et al. A common dominant TLR5 stop codon polymorphism abolishes flagellin signaling and is associated with susceptibility to legionnaires' disease. J. Exp. Med. 198, 1563–1572 (2003).

  9. 9.

    et al. CCR7 is critically important for migration of dendritic cells in intestinal lamina propria to mesenteric lymph nodes. J. Immunol. 176, 803–810 (2006).

  10. 10.

    et al. Salmonella typhimurium translocates flagellin across intestinal epithelia, inducing a proinflammatory response. J. Clin. Invest. 107, 99–109 (2001).

  11. 11.

    , , & Isolation and characterization of antigen-presenting dendritic cells from the mouse intestinal lamina propria. Immunology 70, 40–47 (1990).

  12. 12.

    , , & Dendritic cells: the host Achille's heel for mucosal pathogens? Trends Microbiol. 12, 79–88 (2004).

  13. 13.

    , , , & Phenotypic and functional characterization of CD11c+ dendritic cell population in mouse Peyer's patches. Eur. J. Immunol. 26, 1801–1806 (1996).

  14. 14.

    et al. Characterization of dendritic cells that induce tolerance and T regulatory 1 cell differentiation in vivo. Immunity 18, 605–617 (2003).

  15. 15.

    , , & Operon structure of flagellar genes in Salmonella typhimurium. Mol. Gen. Genet. 214, 11–15 (1988).

  16. 16.

    , & The distribution, ontogeny and origin in the rat of Ia-positive cells with dendritic morphology and of Ia antigen in epithelia, with special reference to the intestine. Eur. J. Immunol. 13, 112–122 (1983).

  17. 17.

    Anatomical basis of tolerance and immunity to intestinal antigens. Nat. Rev. Immunol. 3, 331–341 (2003).

  18. 18.

    , , & Immunomodulatory dendritic cells in intestinal lamina propria. Eur. J. Immunol. 35, 1831–1840 (2005).

  19. 19.

    et al. Oral tolerance originates in the intestinal immune system and relies on antigen carriage by dendritic cells. J. Exp. Med. 203, 519–527 (2006).

  20. 20.

    et al. CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance. Science 307, 254–258 (2005).

  21. 21.

    et al. Evasion of Toll-like receptor 5 by flagellated bacteria. Proc. Natl. Acad. Sci. USA 102, 9247–9252 (2005).

  22. 22.

    , , & A recombinant Salmonella typhimurium vaccine strain is taken up and survives within murine Peyer's patch dendritic cells. Cell. Microbiol. 2, 59–68 (2000).

  23. 23.

    et al. Dendritic cells express tight junction proteins and penetrate gut epithelial monolayers to sample bacteria. Nat. Immunol. 2, 361–367 (2001).

  24. 24.

    , & The functional interface between Salmonella and its host cell: opportunities for therapeutic intervention. Trends Pharmacol. Sci. 26, 564–570 (2005).

  25. 25.

    et al. Extraintestinal dissemination of Salmonella by CD18-expressing phagocytes. Nature 401, 804–808 (1999).

  26. 26.

    et al. Cutting edge: 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).

  27. 27.

    & Plasmid-associated virulence of Salmonella typhimurium. Infect. Immun. 55, 2891–2901 (1987).

  28. 28.

    , , & The roles of Toll-like receptor 9, MyD88, and DNA-dependent protein kinase catalytic subunit in the effects of two distinct CpG DNAs on dendritic cell subsets. J. Immunol. 170, 3059–3064 (2003).

Download references

Acknowledgements

We thank K. Smith and T. Hawn (Institute for Systems Biology, Seattle, Washington) for providing purified flagellin; C. Sasagawa and T. Suzuki (Institute of Medical Science, Tokyo, Japan) for providing bacteria; members of the DNA-chip Development Center for Infectious Diseases (RIMD, Osaka University, Osaka, Japan) for technical advice; N. Kitagaki for technical assistance; and M. Hashimoto for secretarial assistance. Supported by Special Coordination Funds, the Ministry of Education, Culture, Sports, Science and Technology, and Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.

Author information

Author notes

    • Satoshi Uematsu
    •  & Myoung Ho Jang

    These authors contributed equally to this work.

Affiliations

  1. Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Suita Osaka 565-0871, Japan.

    • Satoshi Uematsu
    • , Nicolas Chevrier
    • , Yutaro Kumagai
    • , Masahiro Yamamoto
    • , Hiroki Kato
    • , Hiroaki Hemmi
    • , Osamu Takeuchi
    •  & Shizuo Akira
  2. Laboratory of Immunodynamics, Department of Microbiology and Immunology, Osaka University Graduate School of Medicine (C8), 2-2, Yamada-oka, Suita, 565-0871, Japan.

    • Myoung Ho Jang
    • , Zijin Guo
    • , Nagako Sougawa
    •  & Masayuki Miyasaka
  3. Laboratory of Immunoregulation, Kitasato Institute for Life Sciences and Graduate School of Infection, Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.

    • Hidenori Matsui
  4. Department of Molecular Genetics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

    • Hirotaka Kuwata
    •  & Kiyoshi Takeda
  5. 21st Century COE, Combined Program on Microbiology and Immunology, Osaka University, 3-1 Yamada-oka, Suita Osaka 565-0871, Japan.

    • Cevayir Coban
  6. ERATO, Japan Science and Technology Corporation, 3-1 Yamada-oka, Suita Osaka 565-0871, Japan.

    • Taro Kawai
    • , Ken J Ishii
    • , Osamu Takeuchi
    •  & Shizuo Akira

Authors

  1. Search for Satoshi Uematsu in:

  2. Search for Myoung Ho Jang in:

  3. Search for Nicolas Chevrier in:

  4. Search for Zijin Guo in:

  5. Search for Yutaro Kumagai in:

  6. Search for Masahiro Yamamoto in:

  7. Search for Hiroki Kato in:

  8. Search for Nagako Sougawa in:

  9. Search for Hidenori Matsui in:

  10. Search for Hirotaka Kuwata in:

  11. Search for Hiroaki Hemmi in:

  12. Search for Cevayir Coban in:

  13. Search for Taro Kawai in:

  14. Search for Ken J Ishii in:

  15. Search for Osamu Takeuchi in:

  16. Search for Masayuki Miyasaka in:

  17. Search for Kiyoshi Takeda in:

  18. Search for Shizuo Akira in:

Contributions

S.U. and M.H.J. did most of the experiments to characterize mouse phenotypes; N.C. helped with the quantitative PCR, microarray analysis, isolation of cells and enzyme-linked immunosorbent assays; Z.G. helped to isolate cells and with immunostaining and did the surgical operations for the intestinal loop assay; Y.K. helped with analysis of microarray data; M.Y. helped to generate Tlr5−/− mice; H.K. helped with the enzyme-linked immunosorbent assays; N.S. helped to isolate cells; H.M. provided S. typhimurium and provided instructions for infection experiments; H.K. helped with the infection experiments; H.H. helped to generate Tlr5−/− mice; C.C. helped with the infection experiments; T.K., K.J.I. and O.T. provided advice for the experiments; M.M. provided advice for the experiments and manuscript; K.T. helped to generate Tlr5−/− mice and to design experiments; and S.A. designed all the experiments and prepared the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Shizuo Akira.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Generation of Tlr5−/− mice.

  2. 2.

    Supplementary Fig. 2

    CD11c+ LPCs produce IL-6 in response to TLR2 and TLR9 stimulation.

  3. 3.

    Supplementary Fig. 3

    Surface phenotype of MLN cells 2 d after oral S. typhimurium infection.

  4. 4.

    Supplementary Fig. 4

    Uptake of S. typhimurium in situ.

  5. 5.

    Supplementary Table 1

    Primer sequences.

  6. 6.

    Supplementary Methods

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ni1362

Further reading