Abstract
The innate immune system recognizes pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, but not on the host. Toll-like receptors (TLRs) recognize PAMPs and mediate the production of cytokines necessary for the development of effective immunity1,2,3,4. Flagellin, a principal component of bacterial flagella, is a virulence factor that is recognized by the innate immune system in organisms as diverse as flies, plants and mammals5,6,7,8,9,10,11. Here we report that mammalian TLR5 recognizes bacterial flagellin from both Gram-positive and Gram-negative bacteria, and that activation of the receptor mobilizes the nuclear factor NF-κB and stimulates tumour necrosis factor-α production. TLR5-stimulating activity was purified from Listeria monocytogenes culture supernatants and identified as flagellin by tandem mass spectrometry. Expression of L. monocytogenes flagellin in non-flagellated Escherichia coli conferred on the bacterium the ability to activate TLR5, whereas deletion of the flagellin genes from Salmonella typhimurium abrogated TLR5-stimulating activity. All known TLRs signal through the adaptor protein MyD88. Mice challenged with bacterial flagellin rapidly produced systemic interleukin-6, whereas MyD88-null mice did not respond to flagellin. Our data suggest that TLR5, a member of the evolutionarily conserved Toll-like receptor family, has evolved to permit mammals specifically to detect flagellated bacterial pathogens.
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References
Aderem, A. & Ulevitch, R. J. Toll-like receptors in the induction of the innate immune response. Nature 406, 782–787 (2000).
Brightbill, H. D. & Modlin, R. L. Toll-like receptors: molecular mechanisms of the mammalian immune response. Immunology 101, 1–10 (2000).
Medzhitov, R. & Janeway, C. Jr Innate immune recognition: mechanisms and pathways. Immunol. Rev. 173, 89–97 (2000).
Means, T. K., Golenbock, D. T. & Fenton, M. J. The biology of Toll-like receptors. Cytokine Growth Factor Rev. 11, 219–232 (2000).
Samakovlis, C., Asling, B., Boman, H. G., Gateff, E. & Hultmark, D. In vitro induction of cecropin genes—an immune response in a Drosophila blood cell line. Biochem. Biophys. Res. Commun. 188, 1169–1175 (1992).
Gomez-Gomez, L. & Boller, T. FLS2: an LRR receptor-like kinase involved in the perception of the bacterial elicitor flagellin in Arabidopsis. Mol. Cell 5, 1003–1011 (2000).
Wyant, T. L., Tanner, M. K. & Sztein, M. B. Potent immunoregulatory effects of Salmonella typhi flagella on antigenic stimulation of human peripheral blood mononuclear cells. Infect. Immun. 67, 1338–1346 (1999).
Wyant, T. L., Tanner, M. K. & Sztein, M. B. Salmonella typhi flagella are potent inducers of proinflammatory cytokine secretion by human monocytes. Infect. Immun. 67, 3619–3624 (1999).
Steiner, T. S., Nataro, J. P., Poteet-Smith, C. E., Smith, J. A. & Guerrant, R. L. Enteroaggregative Escherichia coli expresses a novel flagellin that causes IL-8 release from intestinal epithelial cells. J. Clin. Invest. 105, 1769–1777 (2000).
McDermott, P. F., Ciacci-Woolwine, F., Snipes, J. A. & Mizel, S. B. High-affinity interaction between gram-negative flagellin and a cell surface polypeptide results in human monocyte activation. Infect. Immun. 68, 5525–5529 (2000).
Ciacci-Woolwine, F., McDermott, P. F. & Mizel, S. B. Induction of cytokine synthesis by flagella from gram-negative bacteria may be dependent on the activation or differentiation state of human monocytes. Infect. Immun. 67, 5176–5185 (1999).
Medzhitov, R. et al. MyD88 is an adaptor protein in the hToll/IL-1 receptor family signaling pathways. Mol. Cell 2, 253–258 (1998).
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).
Chaudhary, P. M. et al. Cloning and characterization of two Toll/Interleukin-1 receptor-like genes TIL3 and TIL4: evidence for a multi-gene receptor family in humans. Blood 91, 4020–4027 (1998).
Sebastiani, G. 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).
Muzio, M. et al. Differential expression and regulation of toll-like receptors (TLR) in human leukocytes: selective expression of TLR3 in dendritic cells. J. Immunol. 164, 5998–6004 (2000).
Cario, E. & Podolsky, D. K. Differential alteration in intestinal epithelial cell expression of Toll-like receptor 3 (TLR3) and TLR4 in inflammatory bowel disease. Infect. Immun. 68, 7010–7017 (2000).
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).
Ozinsky, A. et al. The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between Toll-like receptors. Proc. Natl Acad. Sci. USA 97, 13766–13771 (2000).
Eng, J. K., McCormack, A. L. & Yates, J. R. An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J. Am. Soc. Mass Spectrom. 5, 976–989 (1994).
Peel, M., Donachie, W. & Shaw, A. Temperature-dependent expression of flagella of Listeria monocytogenes studied by electron microscopy, SDS-PAGE and western blotting. J. Gen. Microbiol. 134, 2171–2178 (1988).
Fujita, H., Yamaguchi, S. & Iino, T. Studies on H-O variants in Salmonella in relation to phase variation. J. Gen. Microbiol. 76, 127–134 (1973).
Underhill, D. M. et al. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature 401, 811–815 (1999).
Brightbill, H. D. et al. Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors. Science 285, 732–736 (1999).
das Gracas Luna, M., Sardella, F. F. & Ferreira, L. C. Salmonella flagellin fused with a linear epitope of colonization factor antigen I (CFA/I) can prime antibody responses against homologous and heterologous fimbriae of enterotoxigenic Escherichia coli. Res. Microbiol. 151, 575–582 (2000).
Stocker, B. A. & Newton, S. M. Immune responses to epitopes inserted in Salmonella flagellin. Int. Rev. Immunol. 11, 167–178 (1994).
Mimori-Kiyosue, Y., Vonderviszt, F. & Namba, K. Locations of terminal segments of flagellin in the filament structure and their roles in polymerization and polymorphism. J. Mol. Biol. 270, 222–237 (1997).
Ibrahim, G. F., Fleet, G. H., Lyons, M. J. & Walker, R. A. Method for the isolation of highly purified Salmonella flagellins. J. Clin. Microbiol. 22, 1040–1044 (1985).
Goodlett, D. R. et al. Protein identification with a single accurate mass of a cysteine-containing peptide and constrained database searching. Anal. Chem. 72, 1112–1118 (2000).
Adachi, O. et al. Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. Immunity 9, 143–150 (1998).
Acknowledgements
We thank D. Portnoy, S. R. Swanzy and K. T. Hughes for reagents and discussions; P. deRoos for advice with chromatography; and H. Harowicz for help with animal studies. F.H. and T.R.H. received support from the HHMI. K.D.S. received support from the NIH. This research was funded by NIH grants to A.A.
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Hayashi, F., Smith, K., Ozinsky, A. et al. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099–1103 (2001). https://doi.org/10.1038/35074106
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DOI: https://doi.org/10.1038/35074106
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