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Subversion of Toll-like receptor signaling by a unique family of bacterial Toll/interleukin-1 receptor domain–containing proteins

Nature Medicine volume 14, pages 399–406 (2008)Cite this article

Abstract

Pathogenic microbes have evolved sophisticated molecular strategies to subvert host defenses. Here we show that virulent bacteria interfere directly with Toll-like receptor (TLR) function by secreting inhibitory homologs of the Toll/interleukin-1 receptor (TIR) domain. Genes encoding TIR domain containing–proteins (Tcps) were identified in Escherichia coli CFT073 (TcpC) and Brucella melitensis (TcpB). We found that TcpC is common in the most virulent uropathogenic E. coli strains and promotes bacterial survival and kidney pathology in vivo. In silico analysis predicted significant tertiary structure homology to the TIR domain of human TLR1, and we show that the Tcps impede TLR signaling through the myeloid differentiation factor 88 (MyD88) adaptor protein, owing to direct binding of Tcps to MyD88. Tcps represent a new class of virulence factors that act by inhibiting TLR- and MyD88-specific signaling, thus suppressing innate immunity and increasing virulence.

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Figure 1: Genomic localization and protein structure of TcpC and TcpB.
Figure 2: Tcps reduce cytokine secretion and increase accumulation of intracellular bacteria.
Figure 3: Tcps impair TLR signaling and interact with MyD88.
Figure 4: TcpC is a virulence factor that promotes bacterial burden in the urinary tract and renal tissue damage.
Figure 5: TcpC is secreted by CFT073 E. coli and is subsequently taken up by host cells.
Figure 6: TNF secretion is reduced by the purified TIR domain of TcpC and PAβN as therapeutic compound.

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References

  1. Iwasaki, A. & Medzhitov, R. Toll-like receptor control of the adaptive immune responses. Nat. Immunol. 5, 987–995 (2004).

    Article  CAS  Google Scholar 

  2. Akira, S., Uematsu, S. & Takeuchi, O. Pathogen recognition and innate immunity. Cell 124, 783–801 (2006).

    Article  CAS  Google Scholar 

  3. Svanborg, C. et al. Uropathogenic Escherichia coli as a model of host-parasite interaction. Curr. Opin. Microbiol. 9, 33–39 (2006).

    Article  CAS  Google Scholar 

  4. Takeuchi, O., Hoshino, K. & Akira, S. Cutting edge: TLR2-deficient and MyD88-deficient mice are highly susceptible to staphylococcus aureus infection. J. Immunol. 165, 5392–5396 (2000).

    Article  CAS  Google Scholar 

  5. Abel, B. et al. Toll-like receptor 4 expression is required to control chronic Mycobacterium tuberculosis infection in mice. J. Immunol. 169, 3155–3162 (2002).

    Article  CAS  Google Scholar 

  6. Torres, D. et al. Toll-like receptor 2 is required for optimal control of Listeria monocytogenes infection. Infect. Immun. 72, 2131–2139 (2004).

    Article  CAS  Google Scholar 

  7. Rodriguez, N. et al. Differential involvement of TLR2 and TLR4 in host survival during pulmonary infection with Chlamydia pneumoniae. Eur. J. Immunol. 36, 1145–1155 (2006).

    Article  CAS  Google Scholar 

  8. Poltorak, A. et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085–2088 (1998).

    Article  CAS  Google Scholar 

  9. Kawai, T. & Akira, S. TLR signaling. Cell Death Differ. 13, 816–825 (2006).

    Article  CAS  Google Scholar 

  10. Burns, K. et al. Inhibition of interleukin 1 receptor/Toll-like receptor signaling through the alternatively spliced, short form of MyD88 is due to its failure to recruit IRAK-4. J. Exp. Med. 197, 263–268 (2003).

    Article  Google Scholar 

  11. Bowie, A. et al. A46R and A52R from vaccinia virus are antagonists of host IL-1 and Toll-like receptor signaling. Proc. Natl. Acad. Sci. USA 97, 10162–10167 (2000).

    Article  CAS  Google Scholar 

  12. Harte, M.T. et al. The poxvirus protein A52R targets Toll-like receptor signaling complexes to suppress host defense. J. Exp. Med. 197, 343–351 (2003).

    Article  CAS  Google Scholar 

  13. Stack, J. et al. Vaccinia virus protein A46R targets multiple Toll-like–interleukin-1 receptor adaptors and contributes to virulence. J. Exp. Med. 201, 1007–1018 (2005).

    Article  CAS  Google Scholar 

  14. Newman, R.M., Salunkhe, P., Godzik, A. & Reed, J.C. Identification and characterization of a novel bacterial virulence factor that shares homology with mammalian Toll/interleukin-1 receptor family proteins. Infect. Immun. 74, 594–601 (2006).

    Article  CAS  Google Scholar 

  15. Radons, J. et al. The interleukin 1 (IL-1) receptor accessory protein Toll/IL-1 receptor domain: analysis of putative interaction sites in vitro mutagenesis and molecular modeling. J. Biol. Chem. 278, 49145–49153 (2003).

    Article  CAS  Google Scholar 

  16. Costa, C.P. et al. Role of chlamydial heat shock protein 60 in the stimulation of innate immune cells by Chlamydia pneumoniae. Eur. J. Immunol. 32, 2460–2470 (2002).

    Article  Google Scholar 

  17. Lafont, F. & van der Goot, F.G. Bacterial invasion via lipid rafts. Cell. Microbiol. 7, 613–620 (2005).

    Article  CAS  Google Scholar 

  18. Pannek, S. et al. Multidrug efflux inhibition in Acinetobacter baumannii: comparison between 1-(1-naphthylmethyl)-piperazine and phenyl-arginine-β-naphthylamide. J. Antimicrob. Chemother. 57, 970–974 (2006).

    Article  CAS  Google Scholar 

  19. Frendeus, B. et al. Escherichia coli P fimbriae utilize the Toll-like receptor 4 pathway for cell activation. Mol. Microbiol. 40, 37–51 (2001).

    Article  CAS  Google Scholar 

  20. Samuelsson, P., Hang, L., Wullt, B., Irjala, H. & Svanborg, C. Toll-like receptor 4 expression and cytokine responses in the human urinary tract mucosa. Infect. Immun. 72, 3179–3186 (2004).

    Article  CAS  Google Scholar 

  21. Gorvel, J.P. & Moreno, E. Brucella intracellular life: from invasion to intracellular replication. Vet. Microbiol. 90, 281–297 (2002).

    Article  CAS  Google Scholar 

  22. Delrue, R.M. et al. Identification of Brucella spp. genes involved in intracellular trafficking. Cell. Microbiol. 3, 487–497 (2001).

    Article  CAS  Google Scholar 

  23. Comerci, D.J., Martinez-Lorenzo, M.J., Sieira, R., Gorvel, J.P. & Ugalde, R.A. Essential role of the VirB machinery in the maturation of the Brucella abortus–containing vacuole. Cell. Microbiol. 3, 159–168 (2001).

    Article  CAS  Google Scholar 

  24. Lapaque, N., Moriyon, I., Moreno, E. & Gorvel, J.P. Brucella lipopolysaccharide acts as a virulence factor. Curr. Opin. Microbiol. 8, 60–66 (2005).

    Article  CAS  Google Scholar 

  25. Sola-Landa, A. et al. A two-component regulatory system playing a critical role in plant pathogens and endosymbionts is present in Brucella abortus and controls cell invasion and virulence. Mol. Microbiol. 29, 125–138 (1998).

    Article  CAS  Google Scholar 

  26. Detilleux, P.G., Deyoe, B.L. & Cheville, N.F. Penetration and intracellular growth of Brucella abortus in nonphagocytic cells in vitro. Infect. Immun. 58, 2320–2328 (1990).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Welch, R.A. et al. Extensive mosaic structure revealed by the complete genome sequence of uropathogenic Escherichia coli. Proc. Natl. Acad. Sci. USA 99, 17020–17024 (2002).

    Article  CAS  Google Scholar 

  28. Kern, W.V. et al. Effect of 1-(1-naphthylmethyl)-piperazine, a novel putative efflux pump inhibitor, on antimicrobial drug susceptibility in clinical isolates of Escherichia coli. J. Antimicrob. Chemother. 57, 339–343 (2006).

    Article  CAS  Google Scholar 

  29. Schumacher, A. et al. Effect of 1-(1-naphthylmethyl)-piperazine, a novel putative efflux pump inhibitor, on antimicrobial drug susceptibility in clinical isolates of Enterobacteriaceae other than Escherichia coli. J. Antimicrob. Chemother. 57, 344–348 (2006).

    Article  CAS  Google Scholar 

  30. Hagberg, L. et al. Contribution of adhesion to bacterial persistence in the mouse urinary tract. Infect. Immun. 40, 265–272 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Bean, M.A., Pees, H., Fogh, J.E., Grabstald, H. & Oettgen, H.F. Cytotoxicity of lymphocytes from patients with cancer of the urinary bladder: detection by a 3- H-proline microcytotoxicity test. Int. J. Cancer 14, 186–197 (1974).

    Article  CAS  Google Scholar 

  32. Datsenko, K.A. & Wanner, B.L. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. USA 97, 6640–6645 (2000).

    Article  CAS  Google Scholar 

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Acknowledgements

S. Schubert (Pettenkofer Institut, München, Germany) provided the human uroepithelial cell line HCV29. This work was supported by SFB576, project B10, Deutsche Forschungsgemeinschaft and the Swedish Medical Research Council (grant numbers 07934, 14577, 14578); The Royal Physiographic Society; The Medical Faculty, Lund University; Network of Excellence Europathogenomics; and The Österlund, Lundberg and Wallenberg Foundations. C.S. was a recipient of the Bristol-Myers Squibb unrestricted grant. We thank G. Häcker for critical reading of the manuscript and C. Kirschning (Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, München, Germany), K. Rückdeschel (Institut für Medizinische Mikrobiologie, Virologie und Hygiene, Hamburg, Germany), F. Schmitz (Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, München, Germany) and S. Akira (Research Institute for Microbial Diseases, Osaka, Japan) for the donation of plasmids. We are grateful to S. Bierl and T. Ertl for construction of expression plasmids and experimental assistance.

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Authors and Affiliations

  1. Institut für Medizinische Mikrobiologie, Immunologie und Hygiene, Technische Universität München, Trogerstrasse 30, München, D-81675, Germany

    Christine Cirl, Susanne Duerr, Nina Wantia, Nuria Rodriguez, Hermann Wagner & Thomas Miethke

  2. Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstrasse 9a, München, D-80336, Germany

    Andreas Wieser & Sören Schubert

  3. Department of Microbiology, Immunology and Glycobiology, Institute of Laboratory Medicine, Lund University, Sölvegatan 23, Lund, 22362, Sweden

    Manisha Yadav, Hans Fischer, Dominik Stappert & Catharina Svanborg

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  1. Christine Cirl
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Contributions

C.C. performed pull-down assays, western blotting, cell culture assays and protein purification. A.W. and S.S. generated the E. coli deletion mutant tcpC::kan. M.Y. performed in vivo experiments. S.D. performed luciferase reporter assays and assisted with confocal microscopy. H.F. and D.S. performed PCR analysis of E. coli strains obtained from human beings. N.W. and N.R. assisted in cell culture assays and provided gene-deficient mice. H.W. participated in writing the manuscript. C.S. supervised in vivo experiments and participated in writing the manuscript. T.M. analyzed the data, designed the whole project and wrote the manuscript.

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Correspondence to Thomas Miethke.

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Cirl, C., Wieser, A., Yadav, M. et al. Subversion of Toll-like receptor signaling by a unique family of bacterial Toll/interleukin-1 receptor domain–containing proteins. Nat Med 14, 399–406 (2008). https://doi.org/10.1038/nm1734

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