Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Aryl hydrocarbon receptor control of a disease tolerance defence pathway

This article has been updated

Abstract

Disease tolerance is the ability of the host to reduce the effect of infection on host fitness. Analysis of disease tolerance pathways could provide new approaches for treating infections and other inflammatory diseases. Typically, an initial exposure to bacterial lipopolysaccharide (LPS) induces a state of refractoriness to further LPS challenge (endotoxin tolerance). We found that a first exposure of mice to LPS activated the ligand-operated transcription factor aryl hydrocarbon receptor (AhR) and the hepatic enzyme tryptophan 2,3-dioxygenase, which provided an activating ligand to the former, to downregulate early inflammatory gene expression. However, on LPS rechallenge, AhR engaged in long-term regulation of systemic inflammation only in the presence of indoleamine 2,3-dioxygenase 1 (IDO1). AhR-complex-associated Src kinase activity promoted IDO1 phosphorylation and signalling ability. The resulting endotoxin-tolerant state was found to protect mice against immunopathology in Gram-negative and Gram-positive infections, pointing to a role for AhR in contributing to host fitness.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Increased susceptibility of Tdo2–/– and Ahr–/– mice to primary LPS challenge.
Figure 2: l-kynurenine (Kyn) is an endogenous AhR ligand.
Figure 3: Absolute requirement for AhR, functional IDO1 and TGF-β in LPS tolerance.
Figure 4: LPS tolerance induces AhR- and Src-dependent IDO1 phosphorylation.
Figure 5: LPS tolerance ameliorates S. Typhimurium infection.
Figure 6: LPS tolerance protects against Streptococcus immunopathology.

Similar content being viewed by others

Change history

  • 09 July 2014

    Affiliation address 9, for author David Gilot, was given incorrectly and has been updated.

References

  1. Fan, H. & Cook, J. A. Molecular mechanisms of endotoxin tolerance. J. Endotoxin Res. 10, 71–84 (2004)

    CAS  PubMed  Google Scholar 

  2. Pena, O. M., Pistolic, J., Raj, D., Fjell, C. D. & Hancock, R. E. Endotoxin tolerance represents a distinctive state of alternative polarization (M2) in human mononuclear cells. J. Immunol. 186, 7243–7254 (2011)

    CAS  PubMed  Google Scholar 

  3. Krausgruber, T. et al. IRF5 promotes inflammatory macrophage polarization and TH1-TH17 responses. Nature Immunol. 12, 231–238 (2011)

    CAS  Google Scholar 

  4. Abdi, K., Singh, N. J. & Matzinger, P. Lipopolysaccharide-activated dendritic cells: “exhausted” or alert and waiting? J. Immunol. 188, 5981–5989 (2012)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Biswas, S. K. & Lopez-Collazo, E. Endotoxin tolerance: new mechanisms, molecules and clinical significance. Trends Immunol. 30, 475–487 (2009)

    CAS  PubMed  Google Scholar 

  6. Park, S. H., Park-Min, K. H., Chen, J., Hu, X. & Ivashkiv, L. B. Tumor necrosis factor induces GSK3 kinase-mediated cross-tolerance to endotoxin in macrophages. Nature Immunol. 12, 607–615 (2011)

    CAS  Google Scholar 

  7. Doreswamy, V. & Peden, D. B. Modulation of asthma by endotoxin. Clin. Exp. Allergy 41, 9–19 (2011)

    CAS  PubMed  Google Scholar 

  8. Stejskalova, L., Dvorak, Z. & Pavek, P. Endogenous and exogenous ligands of aryl hydrocarbon receptor: current state of art. Curr. Drug Metab. 12, 198–212 (2011)

    CAS  PubMed  Google Scholar 

  9. Quintana, F. J. The aryl hydrocarbon receptor: a molecular pathway for the environmental control of the immune response. Immunology 138, 183–189 (2013)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Kimura, A. et al. Aryl hydrocarbon receptor in combination with Stat1 regulates LPS-induced inflammatory responses. J. Exp. Med. 206, 2027–2035 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Nguyen, L. P. & Bradfield, C. A. The search for endogenous activators of the aryl hydrocarbon receptor. Chem. Res. Toxicol. 21, 102–116 (2008)

    CAS  PubMed  Google Scholar 

  12. Murray, M. F. The human indoleamine 2,3-dioxygenase gene and related human genes. Curr. Drug Metab. 8, 197–200 (2007)

    CAS  PubMed  Google Scholar 

  13. Orabona, C. et al. Toward the identification of a tolerogenic signature in IDO-competent dendritic cells. Blood 107, 2846–2854 (2006)

    CAS  PubMed  Google Scholar 

  14. Stone, T. W., Stoy, N. & Darlington, L. G. An expanding range of targets for kynurenine metabolites of tryptophan. Trends Pharmacol. Sci. 34, 136–143 (2013)

    CAS  PubMed  Google Scholar 

  15. Fallarino, F. et al. The combined effects of tryptophan starvation and tryptophan catabolites down-regulate T cell receptor ζ-chain and induce a regulatory phenotype in naive T cells. J. Immunol. 176, 6752–6761 (2006)

    CAS  PubMed  Google Scholar 

  16. Nguyen, N. T. et al. Aryl hydrocarbon receptor negatively regulates dendritic cell immunogenicity via a kynurenine-dependent mechanism. Proc. Natl Acad. Sci. USA 107, 19961–19966 (2010)

    ADS  CAS  PubMed  Google Scholar 

  17. Mezrich, J. D. et al. An interaction between kynurenine and the aryl hydrocarbon receptor can generate regulatory T cells. J. Immunol. 185, 3190–3198 (2010)

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Romani, L. et al. Defective tryptophan catabolism underlies inflammation in mouse chronic granulomatous disease. Nature 451, 211–215 (2008)

    ADS  CAS  PubMed  Google Scholar 

  19. Changsirivathanathamrong, D. et al. Tryptophan metabolism to kynurenine is a potential novel contributor to hypotension in human sepsis. Crit. Care Med. 39, 2678–2683 (2011)

    CAS  PubMed  Google Scholar 

  20. Jung, I. D. et al. Blockade of indoleamine 2,3-dioxygenase protects mice against lipopolysaccharide-induced endotoxin shock. J. Immunol. 182, 3146–3154 (2009)

    CAS  PubMed  Google Scholar 

  21. Sekine, H. et al. Hypersensitivity of aryl hydrocarbon receptor-deficient mice to lipopolysaccharide-induced septic shock. Mol. Cell. Biol. 29, 6391–6400 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Trifari, S., Kaplan, C. D., Tran, E. H., Crellin, N. K. & Spits, H. Identification of a human helper T cell population that has abundant production of interleukin 22 and is distinct from TH-17, TH1 and TH2 cells. Nature Immunol. 10, 864–871 (2009)

    CAS  Google Scholar 

  23. Howard, G. J., Schlezinger, J. J., Hahn, M. E. & Webster, T. F. Generalized concentration addition predicts joint effects of aryl hydrocarbon receptor agonists with partial agonists and competitive antagonists. Environ. Health Perspect. 118, 666–672 (2009)

    PubMed Central  Google Scholar 

  24. Pandini, A. et al. Detection of the TCDD binding-fingerprint within the Ah receptor ligand binding domain by structurally driven mutagenesis and functional analysis. Biochemistry 48, 5972–5983 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Opitz, C. A. et al. An endogenous tumour-promoting ligand of the human aryl hydrocarbon receptor. Nature 478, 197–203 (2011)

    ADS  CAS  PubMed  Google Scholar 

  26. Fallarino, F., Grohmann, U. & Puccetti, P. Indoleamine 2,3-dioxygenase: from catalyst to signaling function. Eur. J. Immunol. 42, 1932–1937 (2012)

    CAS  PubMed  Google Scholar 

  27. De Luca, A. et al. Functional yet balanced reactivity to Candida albicans requires TRIF, MyD88, and IDO-dependent inhibition of Rorc. J. Immunol. 179, 5999–6008 (2007)

    CAS  PubMed  Google Scholar 

  28. Pallotta, M. T. et al. Indoleamine 2,3-dioxygenase is a signaling protein in long-term tolerance by dendritic cells. Nature Immunol. 12, 870–878 (2011)

    CAS  Google Scholar 

  29. Dong, B. et al. FRET analysis of protein tyrosine kinase c-Src activation mediated via aryl hydrocarbon receptor. Biochim. Biophys. Acta 1810, 427–431 (2011)

    CAS  PubMed  Google Scholar 

  30. Randi, A. S. et al. Hexachlorobenzene triggers AhR translocation to the nucleus, c-Src activation and EGFR transactivation in rat liver. Toxicol. Lett. 177, 116–122 (2008)

    CAS  PubMed  Google Scholar 

  31. Backlund, M. & Ingelman-Sundberg, M. Regulation of aryl hydrocarbon receptor signal transduction by protein tyrosine kinases. Cell. Signal. 17, 39–48 (2005)

    CAS  PubMed  Google Scholar 

  32. Thiennimitr, P. et al. Intestinal inflammation allows Salmonella to use ethanolamine to compete with the microbiota. Proc. Natl Acad. Sci. USA 108, 17480–17485 (2011)

    ADS  CAS  PubMed  Google Scholar 

  33. Puliti, M., Uematsu, S., Akira, S., Bistoni, F. & Tissi, L. Toll-like receptor 2 deficiency is associated with enhanced severity of group B streptococcal disease. Infect. Immun. 77, 1524–1531 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Matzinger, P. & Kamala, T. Tissue-based class control: the other side of tolerance. Nature Rev. Immunol. 11, 221–230 (2011)

    CAS  Google Scholar 

  35. Sander, L. E. et al. Hepatic acute-phase proteins control innate immune responses during infection by promoting myeloid-derived suppressor cell function. J. Exp. Med. 207, 1453–1464 (2010)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  36. Romani, L. & Puccetti, P. Protective tolerance to fungi: the role of IL-10 and tryptophan catabolism. Trends Microbiol. 14, 183–189 (2006)

    CAS  PubMed  Google Scholar 

  37. Belladonna, M. L., Orabona, C., Grohmann, U. & Puccetti, P. TGF-β and kynurenines as the key to infectious tolerance. Trends Mol. Med. 15, 41–49 (2009)

    CAS  PubMed  Google Scholar 

  38. Medzhitov, R., Schneider, D. S. & Soares, M. P. Disease tolerance as a defense strategy. Science 335, 936–941 (2012)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  39. Volpi, C. et al. High doses of CpG oligodeoxynucleotides stimulate a tolerogenic TLR9-TRIF pathway. Nature Commun. 4, 1852 (2013)

    ADS  Google Scholar 

  40. Grohmann, U. et al. CTLA-4-Ig regulates tryptophan catabolism in vivo. Nature Immunol. 3, 1097–1101 (2002)

    CAS  Google Scholar 

  41. Fallarino, F. et al. Modulation of tryptophan catabolism by regulatory T cells. Nature Immunol. 4, 1206–1212 (2003)

    CAS  Google Scholar 

  42. Munn, D. H. et al. Prevention of allogeneic fetal rejection by tryptophan catabolism. Science 281, 1191–1193 (1998)

    ADS  CAS  PubMed  Google Scholar 

  43. Samstein, R. M., Josefowicz, S. Z., Arvey, A., Treuting, P. M. & Rudensky, A. Y. Extrathymic generation of regulatory T cells in placental mammals mitigates maternal-fetal conflict. Cell 150, 29–38 (2012)

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Zelante, T. et al. Tryptophan catabolites from microbiota engage aryl hydrocarbon receptor and balance mucosal reactivity via interleukin-22. Immunity 39, 372–385 (2013)

    CAS  Google Scholar 

  45. Zelante, T., Fallarino, F., Bistoni, F., Puccetti, P. & Romani, L. Indoleamine 2,3-dioxygenase in infection: the paradox of an evasive strategy that benefits the host. Microbes Infect. 11, 133–141 (2009)

    CAS  PubMed  Google Scholar 

  46. Chen, W. IDO: more than an enzyme. Nature Immunol. 12, 809–811 (2011)

    CAS  Google Scholar 

  47. Martinon, F., Mayor, A. & Tschopp, J. The inflammasomes: guardians of the body. Annu. Rev. Immunol. 27, 229–265 (2009)

    CAS  PubMed  Google Scholar 

  48. Chambers, M. C. & Schneider, D. S. Balancing resistance and infection tolerance through metabolic means. Proc. Natl Acad. Sci. USA 109, 13886–13887 (2012)

    ADS  CAS  PubMed  Google Scholar 

  49. Orabona, C. et al. SOCS3 drives proteasomal degradation of indoleamine 2,3-dioxygenase (IDO) and antagonizes IDO-dependent tolerogenesis. Proc. Natl Acad. Sci. USA 105, 20828–20833 (2008)

    ADS  CAS  PubMed  Google Scholar 

  50. Denison, M. S., Pandini, A., Nagy, S. R., Baldwin, E. P. & Bonati, L. Ligand binding and activation of the Ah receptor. Chem. Biol. Interact. 141, 3–24 (2002)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by funding from the Italian Association for Cancer Research (AIRC, to P.P.), Fondazione Italiana Sclerosi Multipla Project No. 2010/R/17 (to F.F.), Associazione Umbra Contro il Cancro (to G.S. & M.A.D.F.), Bayer Grants4Target Focus Grant no. 2012-03-0630 (to A.I., F.F. and D.M.), Bayer Early Career Investigator Award (to D.M.), Grant no. R01ES007685 from the US National Institutes of Environmental Health Sciences (to M.S.D.), the Specific Targeted Research Project FUNMETA (to L.R.), and the Italian Ministry of Health in association with Regione dell’Umbria (GR-2008-1138004 to C.O.) We thank D. Fuchs for serum kynurenine determinations. We also thank G. Andrielli for digital art and image editing and G. Ricci for histopathology.

Author information

Authors and Affiliations

Authors

Contributions

A.B. and M.G. designed and conducted all experiments unless otherwise indicated below; M.T.P., D.M. and C.V. analysed IDO and Src phosphorylation; S.B., E.M.C.M., D.P., M.P. and A.I. conducted bioinformatics studies and statistical analysis. A.M. performed homology modelling and docking studies. R.I., T.Z., M.A.D.F., L.R. and L.T. conducted the in vivo studies with Salmonella and GBS. C.V., M.L.B., C.O., G.S., C.B. and R.B. contributed to specific experimental designs; T.V.L., M.P., H.F. and T.N. made possible, and designed, the experiments with TDO2-deficient mice; J.B.D., G.C.P. and R.M. made possible, and designed, the experiments with IDO2-deficient mice; D.G., M.S.D., G.J.G., M.G., B.V., L.B. and U.G. provided conceptual help and reagents throughout experimentation; F.F. designed and supervised all experiments; P.P. supervised the overall study and wrote the manuscript. F.F. and P.P. share senior authorship on this paper.

Corresponding authors

Correspondence to Francesca Fallarino or Paolo Puccetti.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Increased susceptibility to primary LPS challenge in mice treated with a TDO2 inhibitor.

a, Twelve hours before LPS challenge (10 mg per kg), WT mice were treated with vehicle, the IDO1 and IDO2 inhibitor 1-MT (200 mg per kg), or the TDO2 inhibitor 680C91 (10 mg per kg), control groups receiving 1-MT or 680C91 but no LPS. Survival was monitored every 24 h through day 8 of LPS challenge (n = 10 mice per group per experiment, in one out of three). **P < 0.001 (log-rank test). b, Estimation of LD50 (mg per kg) in mice treated with 1.25, 2.5, 5, 10, 20, 40, or 80 mg per kg LPS. n = 10 per group per dose. LD50 values were calculated by curve-fitting (r2 ≥ 0.95) in one experiment representative of two.

Extended Data Figure 2 Lack of endogenous IL-10 increases susceptibility to endotoxaemia.

a, Survival of WT mice exposed to 10 mg per kg LPS in the presence of anti–IL-10 (0.2 mg per mouse daily, for 4 d, commencing 6 h before challenge) or an isotype control. Data are from three independent experiments (mean ± s.d.). b, Survival of WT and Il10–/– mice treated with 10 mg per kg LPS. **P < 0.001 (log-rank test). c, Survival curves of mice of different genotypes challenged with 10 mg per kg LPS, with or without therapeutic subcutaneous IL-10 at 250 ng per mouse, daily, from challenge (day 0) through day 5. *P < 0.05 (IL-10 versus vehicle). The data show that exogenous IL-10 compensates for both the TDO2 and AhR defects at the lower LPS dosage. IL-10 is protective only in TDO2 knockouts when 20 mg per kg LPS is used.

Extended Data Figure 3 Mutation of Gln 377 to Ala in AhR PAS-B domain does not alter receptor half-life, and apparently results in increased TCDD ligand potency.

a, AhR-deficient cDCs were transfected with WT or AhR(Q377A). After 24 h, cells were incubated with cycloheximide (CXM) (10 µg ml−1) and harvested at different times, lysed, and analysed for AhR expression by immunoblotting, using a specific antibody. β-tubulin was used as a loading control. Data are from one experiment of three. b, Ratios (means ± s.d. of three experiments) of WT or AhR(Q377A) to β-tubulin in transfected cDCs at different times of CHX treatment. (No differences by Student’s t-test.)

Extended Data Figure 4 LPS tolerance potentiates IDO1 expression and AhR activation in splenic cDCs.

a, b, Real-time PCR analysis of Ido1 mRNA expression and immunoblot analysis of IDO1 protein in peritoneal exudate macrophages (MΦ) and neutrophils (Neu) (a), as well as in splenic conventional DCs (cDCs) or plasmacytoid DCs (pDCs) (b). Cells were harvested and purified at 24 h (a) or 72 h (b) of LPS rechallenge. For comparison, samples were included from mice on first exposure to 40 mg per kg LPS (unprimed), as opposed to tolerized mice (primed). Data of Ido1 mRNA fold induction are presented as means ± s.d. of three experiments; *P < 0.05 and **P < 0.001, Shapiro test. Immunoblotting data are from one experiment of three. c, Real-time PCR analysis of Ahr and Cyp1a1 transcript expressions in cDCs from the same mice as in b. **P < 0.001, Shapiro test.

Extended Data Figure 5 Absolute requirement for functional AhR, but not TDO2, in LPS tolerance manifestations.

a, Survival curves of WT and LPS-primed (0.5 mg per kg, day 0) WT (prWT) and AhR-deficient (prAhr–/–) mice after a second challenge (on day +7) with 40 mg per kg LPS. Survival was monitored every 24 h through day 8 of LPS challenge. n = 8–10 mice per group per experiment. One experiment of three. *P < 0.05, log-rank test. b, Survival curves of WT and LPS-primed (10 mg per kg, day 0) WT (prWT) and TDO2-deficient (prTdo2–/–) mice after a second challenge (on day +7) with 40 mg per kg LPS. Survival was monitored every 24 h through day 8 of LPS challenge. n = 8–10 mice per group per experiment. One experiment of three. **P < 0.001, log-rank test.

Extended Data Figure 6 Bioinformatic data from myeloid cDCs data sets.

a, Expression changes of tyrosine kinases in LPS-primed myeloid DCs compared to untreated counterparts. b, Log2 fold changes, depicted as mean values and standard errors.

Extended Data Figure 7 LPS tolerance modulates cytokine production and Foxp3 and Rorc transcription in S. Typhimurium infection.

a, IL-6, IL-1β, TNF-α, IL-10, and TGF-β were measured in caecum cell supernatants from LPS-tolerant mice infected with S. enterica Typhimurium. Data are from three independent experiments (means ± s.d.). *P < 0.05 and **P < 0.001 (Student’s t-test). b, RT–PCR expression of Foxp3 and Rorc transcripts in mesenteric lymph node cells from LPS-tolerant, Salmonella-infected mice. Data (mean ± s.d. of three experiments) are presented as normalized transcript expression in the samples relative to normalized transcript expression in control cultures (that is, cells from vehicle-treated mice, in which fold change = 1; dotted line). *P < 0.05; **P < 0.001 (Shapiro test).

Extended Data Figure 8 Ahr–/– and Ido–/– mice are more susceptible than WT mice to S. Typhimurium infection.

a, Naive mice of different genotypes were challenged intragastrically with S. Typhimurium. Mortality data were recorded (**P < 0.001, WT versus all other genotypes; log-rank test). b, Haematoxylin and eosin staining of mouse caeca was performed at 7 days of infection. Scale bars, 50 μm. One of three experiments. c, Transcript expressions of Il17a, Rorc, Il10, and Foxp3 were quantified in mesenteric lymph node cells. Data (mean ± s.d. of three experiments) are presented as normalized transcript expression in the samples relative to normalized transcript expression in cells from uninfected donors, in which fold change = 1. **P < 0.001 (Shapiro test).

Extended Data Figure 9 LPS tolerance modulates Foxp3 and Rorc transcription in GBS infection.

RT–PCR expression of Foxp3 and Rorc transcripts in joint-draining lymph node cells from LPS-tolerant, GBS-infected mice. Data (mean ± s.d. of three experiments) are presented as normalized transcript expression in the samples relative to normalized transcript expression in control cells (that is, cells from vehicle-treated mice, in which fold change = 1; dotted line). **P < 0.001 (Shapiro test).

Extended Data Figure 10 Ahr–/– and Ido–/– mice are more susceptible than WT mice to GBS immunopathology.

a, Naive mice of different genotypes were infected with GBS (107 c.f.u.). Mortality data were recorded (*P < 0.05 and **P < 0.001, WT versus all other genotypes; log-rank test). b, Haematoxylin and eosin staining of joints was performed at 10 days of infection. Scale bars, 100 μm. One of three experiments. c, Transcript expressions of Il17a, Rorc, Il10, and Foxp3 were quantified in joint-draining lymph nodes. Data (mean ± s.d. of three experiments) are presented as normalized transcript expression in the samples relative to normalized transcript expression in cells from uninfected donors, in which fold change = 1. **P < 0.001 (Shapiro test).

Supplementary information

Supplementary Information

This file contains Supplementary Tables 1-2 and Supplementary References. (PDF 95 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bessede, A., Gargaro, M., Pallotta, M. et al. Aryl hydrocarbon receptor control of a disease tolerance defence pathway. Nature 511, 184–190 (2014). https://doi.org/10.1038/nature13323

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature13323

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing