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Biochemical and biophysical research communications2013; 432(2); 256-261; doi: 10.1016/j.bbrc.2013.02.002

Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites.

Abstract: Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explaining these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be targets for dexamethasone regulation. In vitro treatment of equine peripheral blood mononuclear cells (eqPBMC) with LPS decreased transcripts of tlr4 and increased transcription of md2 (myeloid differentiation factor 2) and cd14 (cluster of differentiation 14). Treatment with dexamethasone rescued transcription of tlr4 after LPS inhibition. LPS-induced transcription of md2 was inhibited in the presence of dexamethasone. Dexamethasone alone did not affect transcription of tlr4 and md2.
Copyright © 2013 Elsevier Inc. All rights reserved.
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Publication Date: 2013-02-10 PubMed ID: 23402753PubMed Central: PMC3695733DOI: 10.1016/j.bbrc.2013.02.002Google Scholar: Lookup
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  • Journal Article
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  • N.I.H.
  • Extramural
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study investigates the effect of lipopolysaccharide (LPS) on the transcription of the Toll-like receptor 4 (TLR4) gene, how the drug dexamethasone reverses this effect, and the role of specific binding sites in this process. The study involved in vitro treatment of equine peripheral blood mononuclear cells and noted changes in transcription of TLR4 and other genes.

Background

  • The trigger for the detrimental effects of septic shock, a critical condition caused by infection, is the interaction between Toll-like receptor 4 (TLR4) and lipopolysaccharide (LPS).
  • While septic shock heavily affects horses and humans, the underlying mechanisms remain unclear.

Similarity in LPS-Sensitive Species

  • An analysis of TLR4 gene promoters, elements that control the transcription of the gene, revealed similarities among LPS-sensitive species (human, chimpanzee, and horse) and remarkable differences with LPS-resistant species like mouse and rat.

Dexamethasone and Conserved NFκB Binding Sites

  • Four conserved nuclear factor kappa B (NFκB) binding sites, regulatory sequences where NFκB protein can attach to DNA, were identified in the tlr4 gene promoters and two in the md2 gene promoter sequences. These sites are most probably the targets of dexamethasone regulation.

In Vitro Experiment Results

  • The researchers conducted an in vitro experiment with equine peripheral blood mononuclear cells. Upon LPS treatment, a decrease in tlr4 transcripts (mRNA molecules copied from the DNA of the tlr4 gene) and an increase in transcription of other genes such as md2 and cd14 were observed.
  • Dexamethasone treatment led to a rescuing or reversal of the tlr4 transcription levels after undergoing LPS-induced inhibition. Furthermore, LPS-induced transcription of the md2 gene was inhibited when dexamethasone was present.
  • Dexamethasone treatment alone did not seem to either increase or decrease the transcription levels of tlr4 and md2 genes.

Cite This Article

APA
Bonin CP, Baccarin RY, Nostell K, Nahum LA, Fossum C, de Camargo MM. (2013). Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites. Biochem Biophys Res Commun, 432(2), 256-261. https://doi.org/10.1016/j.bbrc.2013.02.002

Publication

Biochemical and biophysical research communications
ISSN: 1090-2104
NlmUniqueID: 0372516
Country: United States
Language: English
Volume: 432
Issue: 2
Pages: 256-261

Researcher Affiliations

Bonin, Camila P
  • Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, Brazil. mila_bonin@yahoo.com.br
Baccarin, Raquel Y A
    Nostell, Katarina
      Nahum, Laila A
        Fossum, Caroline
          de Camargo, Maristela M

            MeSH Terms

            • Animals
            • Base Sequence
            • Binding Sites / drug effects
            • Cattle
            • Conserved Sequence
            • Dexamethasone / pharmacology
            • Horses
            • Humans
            • Lipopolysaccharide Receptors / genetics
            • Lipopolysaccharides / immunology
            • Lymphocyte Antigen 96 / genetics
            • Mice
            • NF-kappa B / metabolism
            • Pan troglodytes
            • Promoter Regions, Genetic
            • Rats
            • Swine
            • Toll-Like Receptor 4 / genetics
            • Transcription, Genetic / drug effects
            • Transcription, Genetic / immunology

            Grant Funding

            • D43 TW007012 / FIC NIH HHS
            • P50 AI098507 / NIAID NIH HHS
            • D43TW007012 / FIC NIH HHS

            References

            This article includes 31 references
            1. Moore JN, Morris DD. Endotoxemia and septicemia in horses: experimental and clinical correlates.. J. Am. Vet. Med. Assoc. 1992;200:1903–1914.
              pubmed: 1639695
            2. Rangel-Frausto MS, Pittet D, Costigan M, Hwang T, Davis CS, Wenzel RP. The natural history of the systemic inflammatory response syndrome (SIRS). A prospective study.. JAMA 1995;273:117–123.
              pubmed: 7799491
            3. Shuster R, Traub-Dargatz J, Baxter G. Survey of diplomates of the American College of Veterinary Internal Medicine and the American College of Veterinary Surgeons regarding clinical aspects and treatment of endotoxemia in horses.. J. Am. Vet. Med. Assoc. 1997;210:87–92.
              pubmed: 8977655
            4. Suzuki K, Tateda K, Matsumoto T, Gondaira F, Tsujimoto S, Yamaguchi K. Effects of interaction between Escherichia coli verotoxin and lipopolysaccharide on cytokine induction and lethality in mice.. J. Med. Microbiol. 2000;49:905–910.
              pubmed: 11023187
            5. Soop A, Sunden-Cullberg J, Albert J, Hallstrom L, Treutiger CJ, Sollevi A. Adenosine infusion attenuates soluble RAGE in endotoxin-induced inflammation in human volunteers.. Acta Physiol. (Oxf.) 2009;197:47–53.
              pubmed: 19302259
            6. Morris DD, Crowe N, Moore JN. Correlation of clinical and laboratory data with serum tumor necrosis factor activity in horses with experimentally induced endotoxemia.. Am. J. Vet. Res. 1990;51:1935–1940.
              pubmed: 2085219
            7. Schumann RR, Leong SR, Flaggs GW, Gray PW, Wright SD, Mathison JC, Tobias PS, Ulevitch RJ. Structure and function of lipopolysaccharide binding protein.. Science 1990;249:1429–1431.
              pubmed: 2402637
            8. Wright SD, Ramos RA, Tobias PS, Ulevitch RJ, Mathison JC. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein.. Science 1990;249:1431–1433.
              pubmed: 1698311
            9. Poltorak A, He X, Smirnova I, Liu MY, Van Huffel C, Du X, Birdwell D, Alejos E, Silva M, Galanos C, Freudenberg M, Ricciardi-Castagnoli P, Layton B, Beutler B. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene.. Science 1998;282:2085–2088.
              pubmed: 9851930
            10. Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, Miyake K, Kimoto M. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4.. J. Exp. Med. 1999;189:1777–1782.
              pmc: PMC2193086pubmed: 10359581
            11. Hoshino K, Takeuchi O, Kawai T, Sanjo H, Ogawa T, Takeda Y, Takeda K, Akira S. Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product.. J. Immunol. 1999;162:3749–3752.
              pubmed: 10201887
            12. Nomura F, Akashi S, Sakao Y, Sato S, Kawai T, Matsumoto M, Nakanishi K, Kimoto M, Miyake K, Takeda K, Akira S. Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface Toll-like receptor 4 expression.. J. Immunol. 2000;164:3476–3479.
              pubmed: 10725699
            13. Goldring CE, Reveneau S, Pinard D, Jeannin JF. Hyporesponsiveness to lipopolysaccharide alters the composition of NF-kappaB binding to the regulatory regions of inducible nitric oxide synthase gene.. Eur. J. Immunol. 1998;28:2960–2970.
              pubmed: 9754583
            14. Kastenbauer S, Ziegler-Heitbrock HW. NF-kappaB1 (p50) is upregulated in lipopolysaccharide tolerance and can block tumor necrosis factor gene expression.. Infect. Immun. 1999;67:1553–1559.
              pmc: PMC96496pubmed: 10084986
            15. Ziegler-Heitbrock HW, Wedel A, Schraut W, Strobel M, Wendelgass P, Sternsdorf T, Bauerle PA, Haas JG, Riethmuller G. Tolerance to lipopolysaccharide involves mobilization of nuclear factor kappa B with predominance of p50 homodimers.. J. Biol. Chem. 1994;269:17001–17004.
              pubmed: 7516328
            16. Werners AH, Bull S, Vendrig JC, Smyth T, Bosch RR, Fink-Gremmels J, Bryant CE. Genotyping of Toll-like receptor 4, myeloid differentiation factor 2 and CD-14 in the horse: an investigation into the influence of genetic polymorphisms on the LPS induced TNF-alpha response in equine whole blood.. Vet. Immunol. Immunopathol. 2006;111:165–173.
              pubmed: 16476493
            17. Ohto U, Yamakawa N, Akashi-Takamura S, Miyake K, Shimizu T. Structural analyses of human Toll-like receptor 4 polymorphisms D299G and T399I.. J. Biol. Chem. 2012;287:40611–40617.
              pmc: PMC3504774pubmed: 23055527
            18. Hubbard TJ, Aken BL, Ayling S, Ballester B, Beal K, Bragin E, Brent S, Chen Y, Clapham P, Clarke L, Coates G, Fairley S, Fitzgerald S, Fernandez-Banet J, Gordon L, Graf S, Haider S, Hammond M, Holland R, Howe K, Jenkinson A, Johnson N, Kahari A, Keefe D, Keenan S, Kinsella R, Kokocinski F, Kulesha E, Lawson D, Longden I, Megy K, Meidl P, Overduin B, Parker A, Pritchard B, Rios D, Schuster M, Slater G, Smedley D, Spooner W, Spudich G, Trevanion S, Vilella A, Vogel J, White S, Wilder S, Zadissa A, Birney E, Cunningham F, Curwen V, Durbin R, Fernandez-Suarez XM, Herrero J, Kasprzyk A, Proctor G, Smith J, Searle S, Flicek P. Ensembl 2009.. Nucleic Acids Res. 2009;37:D690–697.
              pmc: PMC2686571pubmed: 19033362
            19. Ratnere I, Dubchak I. Obtaining comparative genomic data with the VISTA family of computational tools.. Curr. Protoc. Bioinformatics 2009 Chapter 10 Unit 10 16.
              doi: 10.1002/0471250953.bi1006s26pubmed: 19496056google scholar: lookup
            20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2( Delta Delta C(T)) Method.. Methods 2001;25:402–408.
              pubmed: 11846609
            21. Sherwin RL, Garcia AJ, Bilkovski R. Do low-dose corticosteroids improve mortality or shock reversal in patients with septic shock? A systematic review and position statement prepared for the American Academy of Emergency Medicine.. J. Emerg. Med. 2012;43:7–12.
              pubmed: 22221983
            22. Sligl WI, Milner DA Jr, Sundar S, Mphatswe W, Majumdar SR. Safety and efficacy of corticosteroids for the treatment of septic shock: a systematic review and meta-analysis.. Clin. Infect. Dis. 2009;49:93–101.
              pubmed: 19489712
            23. O'Neill LA. How Toll-like receptors signal: what we know and what we don't know.. Curr. Opin. Immunol. 2006;18:3–9.
              pubmed: 16343886
            24. Banerjee A, Gerondakis S. Coordinating TLR-activated signaling pathways in cells of the immune system.. Immunol. Cell Biol. 2007;85:420–424.
              pubmed: 17637696
            25. Monie TP, Bryant CE, Gay NJ. Activating immunity: lessons from the TLRs and NLRs.. Trends Biochem. Sci. 2009;34:553–561.
              pubmed: 19818630
            26. McCoy CE, Carpenter S, Palsson-McDermott EM, Gearing LJ, O'Neill LA. Glucocorticoids inhibit IRF3 phosphorylation in response to Toll-like receptor-3 and -4 by targeting TBK1 activation.. J. Biol. Chem. 2008;283:14277–14285.
              pubmed: 18356163
            27. Foteinou PT, Calvano SE, Lowry SF, Androulakis IP. In silico simulation of corticosteroids effect on an NFkB-dependent physicochemical model of systemic inflammation.. PLoS One 2009;4:e4706.
              pmc: PMC2651450pubmed: 19274080
            28. Medvedev AE, Henneke P, Schromm A, Lien E, Ingalls R, Fenton MJ, Golenbock DT, Vogel SN. Induction of tolerance to lipopolysaccharide and mycobacterial components in Chinese hamster ovary/CD14 cells is not affected by overexpression of Toll-like receptors 2 or 4.. J. Immunol. 2001;167:2257–2267.
              pubmed: 11490013
            29. Tissieres P, Dunn-Siegrist I, Schappi M, Elson G, Comte R, Nobre V, Pugin J. Soluble MD-2 is an acute-phase protein and an opsonin for gram-negative bacteria.. Blood 2008;111:2122–2131.
              pubmed: 18056837
            30. Yan Q, Carmody RJ, Qu Z, Ruan Q, Jager J, Mullican SE, Lazar MA, Chen YH. Nuclear factor-kappa B binding motifs specify Toll-like receptor-induced gene repression through an inducible repressosome.. Proc. Natl. Acad. Sci. USA 2012;109:14140–14145.
              pmc: PMC3435186pubmed: 22891325
            31. Bolker J. Model organisms: there's more to life than rats and flies.. Nature 2012;491:31–33.
              pubmed: 23128209

            Citations

            This article has been cited 5 times.
            1. Mahmoud J, Bovy MA, Heming N, Annane D. Corticosteroids in sepsis. J Intensive Med 2025 Oct;5(4):304-312.
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            2. Hsieh WT, Hsu MH, Lin WJ, Xiao YC, Lyu PC, Liu YC, Lin WY, Kuo YH, Chung JG. Ergosta-7, 9 (11), 22-trien-3β-ol Interferes with LPS Docking to LBP, CD14, and TLR4/MD-2 Co-Receptors to Attenuate the NF-κB Inflammatory Pathway In Vitro and Drosophila. Int J Mol Sci 2021 Jun 17;22(12).
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            3. Cao W, Huang H, Xia T, Liu C, Muhammad S, Sun C. Homeobox a5 Promotes White Adipose Tissue Browning Through Inhibition of the Tenascin C/Toll-Like Receptor 4/Nuclear Factor Kappa B Inflammatory Signaling in Mice. Front Immunol 2018;9:647.
              doi: 10.3389/fimmu.2018.00647pubmed: 29651293google scholar: lookup
            4. Goodwin JE, Feng Y, Velazquez H, Zhou H, Sessa WC. Loss of the endothelial glucocorticoid receptor prevents the therapeutic protection afforded by dexamethasone after LPS. PLoS One 2014;9(10):e108126.
              doi: 10.1371/journal.pone.0108126pubmed: 25299055google scholar: lookup
            5. Haim YO, Unger ND, Souroujon MC, Mittelman M, Neumann D. Resistance of LPS-activated bone marrow derived macrophages to apoptosis mediated by dexamethasone. Sci Rep 2014 Mar 10;4:4323.
              doi: 10.1038/srep04323pubmed: 24608810google scholar: lookup
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