Skip to main content

Advertisement

SpringerLink
Log in

Bacterial Cell Wall Constituents Induce Hepcidin Expression in Macrophages Through MyD88 Signaling

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Hepcidin is a key regulator of iron recycling by macrophages that is synthesized mainly by hepatocytes but also by macrophages. However, very little is known about the molecular regulation of hepcidin in macrophages. In the present study, we investigated hepcidin regulation in the RAW264.7 macrophage cell line and in murine peritoneal macrophages stimulated with different Toll-like receptor (TLR) ligands. We found that TLR-2 and TLR-4 ligands activated hepcidin expression in RAW264.7 cells and in wild-type murine peritoneal macrophages, but not in murine peritoneal macrophages isolated from TLR2−/−, TLR-4-deficient or MyD88−/− mice. IL-6 production by RAW264.7 cells stimulated with lipopolysaccharide (LPS, TLR4 ligand) was enhanced by high amounts of iron present in the culture medium. We conclude that hepcidin expression in macrophages is regulated mainly through TLR2 and TLR4 receptors via the MyD88-dependent signaling pathway and that autocrine regulation of iron accumulation in macrophages by hepcidin may affect the levels of proinflammatory cytokine production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Schaible, U.E., and S.H.E. Kaufmann. 2004. Iron and microbial infection. Nat Rev Micro 2: 946–953.

    Article  CAS  Google Scholar 

  2. T. Nevitt, War-Fe-re: iron at the core of fungal virulence and host immunity. Biometals 24: 547-58.

  3. Collins, H.L. 2008. Withholding iron as a cellular defence mechanism—friend or foe? European Journal of Immunology 38: 1803–6.

    Article  PubMed  CAS  Google Scholar 

  4. Ganz, T., and E. Nemeth. 2009. Iron sequestration and anemia of inflammation. Sem Hemat 46: 387–393.

    Article  CAS  Google Scholar 

  5. Ganz, T. 2005. Hepcidin—a regulator of intestinal iron absorption and iron recycling by macrophages. Best Practice & Research. Clinical Haematology 18: 171–82.

    Article  CAS  Google Scholar 

  6. Nemeth, E., M.S. Tuttle, J. Powelson, M.B. Vaughn, A. Donovan, D.M. Ward, T. Ganz, and J. Kaplan. 2004. Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization. Science 306: 2090–3.

    Article  PubMed  CAS  Google Scholar 

  7. Constante, M., W. Jiang, D. Wang, V.-A. Raymond, M. Bilodeau, and M.M. Santos. 2006. Distinct requirements for Hfe in basal and induced hepcidin levels in iron overload and inflammation. American Journal of Physiology. Gastrointestinal and Liver Physiology 291: G229–237.

    Article  PubMed  CAS  Google Scholar 

  8. Nicolas, G., C. Chauvet, L. Viatte, J.L. Danan, X. Bigard, I. Devaux, C. Beaumont, A. Kahn, and S. Vaulont. 2002. The gene encoding the iron regulatory peptide hepcidin is regulated by anemia, hypoxia, and inflammation. The Journal of Clinical Investigation 110: 1037–44.

    PubMed  CAS  Google Scholar 

  9. Weiss, G., and L.T. Goodnough. 2005. Anemia of chronic disease. The New England Journal of Medicine 352: 1011–1023.

    Article  PubMed  CAS  Google Scholar 

  10. Nguyen, N.B., K.D. Callaghan, A.J. Ghio, D.J. Haile, and F. Yang. 2006. Hepcidin expression and iron transport in alveolar macrophages. American Journal of Physiology. Lung Cellular and Molecular Physiology 291: L417–L425.

    Article  PubMed  CAS  Google Scholar 

  11. Sow, F.B., W.C. Florence, A.R. Satoskar, L.S. Schlesinger, B.S. Zwilling, and W.P. Lafuse. 2007. Expression and localization of hepcidin in macrophages: a role in host defense against tuberculosis. Journal of Leukocyte Biology 82: 934–45.

    Article  PubMed  CAS  Google Scholar 

  12. Theurl, I., M. Theurl, M. Seifert, S. Mair, M. Nairz, H. Rumpold, H. Zoller, R. Bellmann-Weiler, H. Niederegger, H. Talasz, and G. Weiss. 2008. Autocrine formation of hepcidin induces iron retention in human monocytes. Blood 111: 2392–2399.

    Article  PubMed  CAS  Google Scholar 

  13. Kumar, H., T. Kawai, and S. Akira. 2011. Pathogen recognition by the innate immune system. International Reviews of Immunology 30: 16–34.

    Article  PubMed  CAS  Google Scholar 

  14. Akira, S., S. Uematsu, and O. Takeuchi. 2006. Pathogen recognition and innate immunity. Cell 124: 783–801.

    Article  PubMed  CAS  Google Scholar 

  15. Lee, P., H. Peng, T. Gelbart, L. Wang, and E. Beutler. 2005. Regulation of hepcidin transcription by interleukin-1 and interleukin-6. Proc Natl Acad Sci USA 102: 1906–10.

    Article  PubMed  CAS  Google Scholar 

  16. Akira, S., K. Hoshino, and T. Kaisho. 2000. The role of Toll-like receptors and MyD88 in innate immune responses. Journal of Endotoxin Research 6: 383–7.

    PubMed  CAS  Google Scholar 

  17. Miranda, C.J., H. Makui, N.C. Andrews, and M.M. Santos. 2004. Contributions of beta2-microglobulin-dependent molecules and lymphocytes to iron regulation: insights from HfeRag1(−/−) and beta2mRag1(−/−) double knock-out mice. Blood 103: 2847–9.

    Article  PubMed  CAS  Google Scholar 

  18. Yu, Y., and D.R. Richardson. 2011. Cellular iron depletion stimulates the JNK and p38 MAPK signaling transduction pathways, dissociation of ASK1-thioredoxin, and activation of ASK1. Journal of Biological Chemistry 286: 15413–15427.

    Article  PubMed  CAS  Google Scholar 

  19. Coppola, G., S.-H. Choi, M.M. Santos, C.J. Miranda, D. Tentler, E.M. Wexler, M. Pandolfo, and D.H. Geschwind. 2006. Gene expression profiling in frataxin deficient mice: microarray evidence for significant expression changes without detectable neurodegeneration. Neurobiology of Disease 22: 302–311.

    Article  PubMed  CAS  Google Scholar 

  20. Jiang, W., M. Constante, and M.M. Santos. 2008. Anemia upregulates lipocalin 2 in the liver and serum. Blood Cells, Molecules & Diseases 41: 169–174.

    Article  CAS  Google Scholar 

  21. Constante, M., D. Wang, V.A. Raymond, M. Bilodeau, and M.M. Santos. 2007. Repression of repulsive guidance molecule C during inflammation is independent of Hfe and involves tumor necrosis factor-alpha. American Journal of Pathology 170: 497–504.

    Article  PubMed  CAS  Google Scholar 

  22. Nhu, Q.M., N. Cuesta, and S.N. Vogel. 2006. Transcriptional regulation of lipopolysaccharide (LPS)-induced Toll-like receptor (TLR) expression in murine macrophages: role of interferon regulatory factors 1 (IRF-1) and 2 (IRF-2). Journal of Endotoxin Research 12: 285–295.

    PubMed  CAS  Google Scholar 

  23. Qureshi, S.T., L. Larivière, G. Leveque, S. Clermont, K.J. Moore, P. Gros, and D. Malo. 1999. Endotoxin-tolerant mice have mutations in toll-like receptor 4 (Tlr4). The Journal of Experimental Medicine 189: 615–625.

    Article  PubMed  CAS  Google Scholar 

  24. Poltorak, A., X. He, I. Smirnova, M.-Y. Liu, C.V. Huffel, X. Du, D. Birdwell, E. Alejos, M. Silva, C. Galanos, M. Freudenberg, P. Ricciardi-Castagnoli, B. Layton, and B. Beutler. 1998. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282: 2085–2088.

    Article  PubMed  CAS  Google Scholar 

  25. Nemeth, E., S. Rivera, V. Gabayan, C. Keller, S. Taudorf, B.K. Pedersen, and T. Ganz. 2004. IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin. The Journal of Clinical Investigation 113: 1271–6.

    PubMed  CAS  Google Scholar 

  26. Peyssonnaux, C., A.S. Zinkernagel, V. Datta, X. Lauth, R.S. Johnson, and V. Nizet. 2006. TLR4-dependent hepcidin expression by myeloid cells in response to bacterial pathogens. Blood 107: 3727–32.

    Article  PubMed  CAS  Google Scholar 

  27. Nemeth, E., E.V. Valore, M. Territo, G. Schiller, A. Lichtenstein, and T. Ganz. 2003. Hepcidin, a putative mediator of anemia of inflammation, is a type II acute-phase protein. Blood 101: 2461–3.

    Article  PubMed  CAS  Google Scholar 

  28. Lee, P., H. Peng, T. Gelbart, and E. Beutler. 2004. The IL-6- and lipopolysaccharide-induced transcription of hepcidin in HFE-, transferrin receptor 2-, and beta 2-microglobulin-deficient hepatocytes. Proc Natl Acad Sci USA 101: 9263–5.

    Article  PubMed  CAS  Google Scholar 

  29. Chaston, T., B. Chung, M. Mascarenhas, J. Marks, B. Patel, S.K. Srai, and P. Sharp. 2008. Evidence for differential effects of hepcidin in macrophages and intestinal epithelial cells. Gut 57: 374–382.

    Article  PubMed  CAS  Google Scholar 

  30. Krause, A., S. Neitz, H.-J. Magert, A. Schulz, W.-G. Forssmann, P. Schulz-Knappe, and K. Adermann. 2000. LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial activity. FEBS Letters 480: 147–150.

    Article  PubMed  CAS  Google Scholar 

  31. Barthe, C., A. Hocquellet, and B. Garbay. 2011. Bacteriostatic activity of the proregion of human hepcidin. Protein and Peptide Letters 18: 36–40.

    Article  PubMed  CAS  Google Scholar 

  32. Levy, J.E., L.K. Montross, and N.C. Andrews. 2000. Genes that modify the hemochromatosis phenotype in mice. The Journal of Clinical Investigation 105: 1209–16.

    Article  PubMed  CAS  Google Scholar 

  33. Allen, K.J., L.C. Gurrin, C.C. Constantine, N.J. Osborne, M.B. Delatycki, A.J. Nicoll, C.E. McLaren, M. Bahlo, A.E. Nisselle, C.D. Vulpe, G.J. Anderson, M.C. Southey, G.G. Giles, D.R. English, J.L. Hopper, J.K. Olynyk, L.W. Powell, and D.M. Gertig. 2008. Iron-overload-related disease in HFE hereditary hemochromatosis. The New England Journal of Medicine 358: 221–230.

    Article  PubMed  CAS  Google Scholar 

  34. Wang, L., L. Harrington, E. Trebicka, H.N. Shi, J.C. Kagan, C.C. Hong, H.Y. Lin, J.L. Babitt, and B.J. Cherayil. 2009. Selective modulation of TLR4-activated inflammatory responses by altered iron homeostasis in mice. The Journal of Clinical Investigation 119: 3322–8.

    PubMed  CAS  Google Scholar 

  35. Wang, L., E.E. Johnson, H.N. Shi, W.A. Walker, M. Wessling-Resnick, and B.J. Cherayil. 2008. Attenuated inflammatory responses in hemochromatosis reveal a role for iron in the regulation of macrophage cytokine translation. Journal of Immunology 181: 2723–2731.

    CAS  Google Scholar 

Download references

Acknowledgments

We acknowledge the help of Dr. Peter P. Liu from the Heart and Stroke/Richard Lewar Centre of Excellence, University of Toronto, Canada with the MyD88−/− mouse. This work was supported by a grant from the Canadian Institutes of Health Research (CIHR, grant no. MOP44045). AL is the recipient of a Ph.D. scholarship from the Natural Sciences and Engineering Research Council of Canada (NSERC). MMS is the recipient of a research scholarship (Senior) from the FRSQ (Fonds de la recherche en santé du Québec).

Contributions

AL designed and performed experiments, collected, and analyzed the data, and wrote the manuscript; and MMS designed the experiments, analyzed the data, and critically reviewed the manuscript.

Conflict of interest

The authors declare no competing financial interests.

Author information

Authors and Affiliations

  1. Centre de recherche du Centre hospitalier de l’Université de Montréal (CRCHUM), Hôpital Notre-Dame, Pav. De Sève Y5625, 1560 rue Sherbrooke est, Montréal, Québec, H2L 4M1, Canada

    Antonio Layoun

  2. Institut du cancer de Montréal, Montreal, Quebec, Canada

    Antonio Layoun & Manuela M. Santos

  3. Département de Médecine, Université de Montréal, Montréal, Québec, Canada

    Manuela M. Santos

  4. Centre de recherche du Centre hospitalier de l’Université de Montréal (CRCHUM), Pav. De Sève Y5625, 1560 rue Sherbrooke est, Montréal, Québec, H2L 4M1, Canada

    Manuela M. Santos

Authors
  1. Antonio Layoun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Manuela M. Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Manuela M. Santos.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Layoun, A., Santos, M.M. Bacterial Cell Wall Constituents Induce Hepcidin Expression in Macrophages Through MyD88 Signaling. Inflammation 35, 1500–1506 (2012). https://doi.org/10.1007/s10753-012-9463-4

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-012-9463-4

KEY WORDS

Search

Navigation