Massachusetts Biologic Laboratories
Adaptor Proteins, Signal Transducing; Antigens, Differentiation; Bacterial Proteins; Cell Line, Tumor; Cells, Cultured; Central Nervous System Neoplasms; DNA; Enzyme Activation; Epithelial Cells; Flagellin; Glioblastoma; HT29 Cells; HeLa Cells; Humans; I-kappa B Kinase; Inflammation; Intestines; Kidney; Lung Neoplasms; Membrane Glycoproteins; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Myeloid Differentiation Factor 88; NF-kappa B; Protein Binding; Protein-Serine-Threonine Kinases; Receptors, Cell Surface; Receptors, Immunologic; Salmonella; Salmonella Infections; Signal Transduction; Solubility; Toll-Like Receptor 5; Toll-Like Receptors
Life Sciences | Medicine and Health Sciences | Microbiology
BACKGROUND: Infection of intestinal epithelial cells by pathogenic Salmonella leads to activation of signaling cascades that ultimately initiate the proinflammatory gene program. The transcription factor NF-kappa B is a key regulator/activator of this gene program and is potently activated. We explored the mechanism by which Salmonella activates NF-kappa B during infection of cultured intestinal epithelial cells and found that flagellin produced by the bacteria and contained on them leads to NF-kappa B activation in all the cells; invasion of cells by the bacteria is not required to activate NF-kappa B.
RESULTS: Purified flagellin activated the mitogen activated protein kinase (MAPK), stress-activated protein kinase (SAPK) and I kappa B kinase (IKK) signaling pathways that lead to expression of the proinflammatory gene program in a temporal fashion nearly identical to that of infection of intestinal epithelial cells by Salmonella. Flagellin expression was required for Salmonella invasion of host cells and it activated NF-kappa B via toll-like receptor 5 (TLR5). Surprisingly, a number of cell lines found to be unresponsive to flagellin express TLR5 and expression of exogenous TLR5 in these cells induces NF-kappa B activity in response to flagellin challenge although not robustly. Conversely, overexpression of dominant-negative TLR5 alleles only partially blocks NF-kappa B activation by flagellin. These observations are consistent with the possibility of either a very stable TLR5 signaling complex, the existence of a low abundance flagellin co-receptor or required adapter, or both.
CONCLUSION: These collective results provide the evidence that flagellin acts as the main determinant of Salmonella mediated NF-kappa B and proinflammatory signaling and gene activation by this flagellated pathogen. In addition, expression of the fli C gene appears to play an important role in the proper functioning of the TTSS since mutants that fail to express fli C are defective in expressing a subset of Sip proteins and fail to invade host cells. Flagellin added in trans cannot restore the ability of the fli C mutant bacteria to invade intestinal epithelial cells. Lastly, TLR5 expression in weak and non-responding cells indicates that additional factors may be required for efficient signal propagation in response to flagellin recognition.
Rights and Permissions
© 2004 Tallant et al; licensee BioMed Central Ltd. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
DOI of Published Version
BMC Microbiol. 2004 Aug 23;4:33. Link to article on publisher's site
Tallant T, Deb A, Kar N, Lupica J, de Veer MJ, DiDonato JA. (2004). Flagellin Acting Via TLR5 is the Major Activator of Key Signaling Pathways Leading to NF-kappa B and Proinflammatory Gene Program activation in intestinal epithelial cells. Open Access Publications by UMMS Authors. https://doi.org/10.1186/1471-2180-4-33. Retrieved from https://escholarship.umassmed.edu/oapubs/2291