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The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5

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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Figure 1: Constitutively active TLR5 activates NF-κB and TNF-α production.
Figure 2: Bacterial culture supernatants contain TLR5-stimulating activity.
Figure 3: Purification of TLR5-stimulating activity from L. monocytogenes culture supernatant.
Figure 4: Identification by mass spectrometry of flagellin as the TLR5 stimulus.
Figure 5: Flagellin expression in bacteria reconstitutes TLR5-stimulating activity, and deletion of flagellin genes abrogates TLR5-stimulating activity.
Figure 6: Flagellin-induced systemic IL-6 production is dependent on the TLR signalling adaptor molecule, MyD88.

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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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Correspondence to Alan Aderem.

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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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