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:

CD36 coordinates NLRP3 inflammasome activation by facilitating intracellular nucleation of soluble ligands into particulate ligands in sterile inflammation

Nature Immunology volume 14, pages 812–820 (2013)Cite this article

Subjects

Abstract

Particulate ligands, including cholesterol crystals and amyloid fibrils, induce production of interleukin 1β (IL-1β) dependent on the cytoplasmic sensor NLRP3 in atherosclerosis, Alzheimer's disease and diabetes. Soluble endogenous ligands, including oxidized low-density lipoprotein (LDL), amyloid-β and amylin peptides, accumulate in such diseases. Here we identify an endocytic pathway mediated by the pattern-recognition receptor CD36 that coordinated the intracellular conversion of those soluble ligands into crystals or fibrils, which resulted in lysosomal disruption and activation of the NLRP3 inflammasome. Consequently, macrophages that lacked CD36 failed to elicit IL-1β production in response to those ligands, and targeting CD36 in atherosclerotic mice resulted in lower serum concentrations of IL-1β and accumulation of cholesterol crystals in plaques. Collectively, our findings highlight the importance of CD36 in the accrual and nucleation of NLRP3 ligands from within the macrophage and position CD36 as a central regulator of inflammasome activation in sterile inflammation.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: CD36-mediated uptake of oxLDL generates intracellular crystals and activates the NLRP3 inflammasome.
Figure 2: CD36-mediated formation of cholesterol crystals and activation of NLRP3 occurs via a lysosomal pathway.
Figure 3: CD36 regulates priming of the NLRP3 inflammasome by oxLDL via TLR4-TLR6.
Figure 4: Inflammasome activity is impaired in atherosclerosis-susceptible mice deficient in CD36 and its signaling partners TLR4 and TLR6.
Figure 5: Uptake of soluble amyloid-β by CD36 induces intracellular NLRP3-activating formation of amyloid.
Figure 6: CD36 regulates the activation of NLRP3 by IAPP.

Similar content being viewed by others

References

  1. Abela, G.S. Cholesterol crystals piercing the arterial plaque and intima trigger local and systemic inflammation. J. Clin. Lipidol. 4, 156–164 (2010).

    Article  PubMed  Google Scholar 

  2. Meyer-Luehmann, M. et al. Rapid appearance and local toxicity of amyloid-β plaques in a mouse model of Alzheimer's disease. Nature 451, 720–724 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Westermark, G.T., Westermark, P., Berne, C. & Korsgren, O. Nordic Network for Clinical Islet T. Widespread amyloid deposition in transplanted human pancreatic islets. N. Engl. J. Med. 359, 977–979 (2008).

    Article  CAS  PubMed  Google Scholar 

  4. Duewell, P. et al. NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Nature 464, 1357–1361 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Halle, A. et al. The NALP3 inflammasome is involved in the innate immune response to amyloid-beta. Nat. Immunol. 9, 857–865 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Masters, S.L. et al. Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1β in type 2 diabetes. Nat. Immunol. 11, 897–904 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Rajamäki, K. et al. Cholesterol crystals activate the NLRP3 inflammasome in human macrophages: a novel link between cholesterol metabolism and inflammation. PLoS ONE 5, e11765 (2010).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Kuida, K. et al. Altered cytokine export and apoptosis in mice deficient in interleukin-1β converting enzyme. Science 267, 2000–2003 (1995).

    Article  CAS  PubMed  Google Scholar 

  9. Keller, M., Ruegg, A., Werner, S. & Beer, H.D. Active caspase-1 is a regulator of unconventional protein secretion. Cell 132, 818–831 (2008).

    Article  CAS  PubMed  Google Scholar 

  10. Bauernfeind, F. et al. Reactive oxygen species inhibitors block priming, but not activation, of the NLRP3 inflammasome. J. Immunol. 187, 613–617 (2011).

    Article  CAS  PubMed  Google Scholar 

  11. Bauernfeind, F.G. et al. NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J. Immunol. 183, 787–791 (2009).

    Article  CAS  PubMed  Google Scholar 

  12. Embry, C.A., Franchi, L., Nunez, G. & Mitchell, T.C. Mechanism of impaired NLRP3 inflammasome priming by monophosphoryl lipid A. Sci. Signal. 4, ra28 (2011).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Hornung, V. et al. Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization. Nat. Immunol. 9, 847–856 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Zhou, R., Yazdi, A.S., Menu, P. & Tschopp, J. A role for mitochondria in NLRP3 inflammasome activation. Nature 469, 221–225 (2011).

    Article  CAS  PubMed  Google Scholar 

  15. Williams, K.J. & Tabas, I. The response-to-retention hypothesis of early atherogenesis. Arterioscler. Thromb. Vasc. Biol. 15, 551–561 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Lim, R.S. et al. Identification of cholesterol crystals in plaques of atherosclerotic mice using hyperspectral CARS imaging. J. Lipid Res. 52, 2177–2186 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Haass, C. et al. Amyloid β-peptide is produced by cultured cells during normal metabolism. Nature 359, 322–325 (1992).

    Article  CAS  PubMed  Google Scholar 

  18. Kirschner, D.A. et al. Synthetic peptide homologous to β protein from Alzheimer disease forms amyloid-like fibrils in vitro. Proc. Natl. Acad. Sci. USA 84, 6953–6957 (1987).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Seubert, P. et al. Isolation and quantification of soluble Alzheimer's β-peptide from biological fluids. Nature 359, 325–327 (1992).

    Article  CAS  PubMed  Google Scholar 

  20. Clark, A. et al. Islet amyloid formed from diabetes-associated peptide may be pathogenic in type-2 diabetes. Lancet 2, 231–234 (1987).

    Article  CAS  PubMed  Google Scholar 

  21. Cooper, G.J. et al. Purification and characterization of a peptide from amyloid-rich pancreases of type 2 diabetic patients. Proc. Natl. Acad. Sci. USA 84, 8628–8632 (1987).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Westermark, P. et al. Amyloid fibrils in human insulinoma and islets of Langerhans of the diabetic cat are derived from a neuropeptide-like protein also present in normal islet cells. Proc. Natl. Acad. Sci. USA 84, 3881–3885 (1987).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Badman, M.K., Pryce, R.A., Charge, S.B., Morris, J.F. & Clark, A. Fibrillar islet amyloid polypeptide (amylin) is internalised by macrophages but resists proteolytic degradation. Cell Tissue Res. 291, 285–294 (1998).

    Article  CAS  PubMed  Google Scholar 

  24. de Koning, E.J. et al. Macrophages and pancreatic islet amyloidosis. Amyloid 5, 247–254 (1998).

    Article  CAS  PubMed  Google Scholar 

  25. Haass, C., Koo, E.H., Mellon, A., Hung, A.Y. & Selkoe, D.J. Targeting of cell-surface β-amyloid precursor protein to lysosomes: alternative processing into amyloid-bearing fragments. Nature 357, 500–503 (1992).

    Article  CAS  PubMed  Google Scholar 

  26. Hartmann, T. et al. Distinct sites of intracellular production for Alzheimer's disease Aβ40/42 amyloid peptides. Nat. Med. 3, 1016–1020 (1997).

    Article  CAS  PubMed  Google Scholar 

  27. Silverstein, R.L. & Febbraio, M. CD36, a scavenger receptor involved in immunity, metabolism, angiogenesis, and behavior. Sci. Signal. 2, re3 (2009).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Kunjathoor, V.V. et al. Scavenger receptors class A-I/II and CD36 are the principal receptors responsible for the uptake of modified low density lipoprotein leading to lipid loading in macrophages. J. Biol. Chem. 277, 49982–49988 (2002).

    Article  CAS  PubMed  Google Scholar 

  29. Moore, K.J. et al. A CD36-initiated signaling cascade mediates inflammatory effects of β-amyloid. J. Biol. Chem. 277, 47373–47379 (2002).

    Article  CAS  PubMed  Google Scholar 

  30. Wilkinson, K., Boyd, J.D., Glicksman, M., Moore, K.J. & El Khoury, J. A high content drug screen identifies ursolic acid as an inhibitor of amyloid beta protein interactions with its receptor CD36. J. Biol. Chem. 286, 34914–34922 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Stewart, C.R. et al. CD36 ligands promote sterile inflammation through assembly of a Toll-like receptor 4 and 6 heterodimer. Nat. Immunol. 11, 155–161 (2010).

    Article  CAS  PubMed  Google Scholar 

  32. El Khoury, J.B. et al. CD36 mediates the innate host response to β-amyloid. J. Exp. Med. 197, 1657–1666 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Febbraio, M. et al. Targeted disruption of the class B scavenger receptor CD36 protects against atherosclerotic lesion development in mice. J. Clin. Invest. 105, 1049–1056 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Moore, K.J. et al. Loss of receptor-mediated lipid uptake via scavenger receptor A or CD36 pathways does not ameliorate atherosclerosis in hyperlipidemic mice. J. Clin. Invest. 115, 2192–2201 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Goudriaan, J.R. et al. CD36 deficiency increases insulin sensitivity in muscle, but induces insulin resistance in the liver in mice. J. Lipid Res. 44, 2270–2277 (2003).

    Article  CAS  PubMed  Google Scholar 

  36. Hajri, T., Han, X.X., Bonen, A. & Abumrad, N.A. Defective fatty acid uptake modulates insulin responsiveness and metabolic responses to diet in CD36-null mice. J. Clin. Invest. 109, 1381–1389 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Kennedy, D.J. et al. A CD36-dependent pathway enhances macrophage and adipose tissue inflammation and impairs insulin signalling. Cardiovasc. Res. 89, 604–613 (2011).

    Article  CAS  PubMed  Google Scholar 

  38. Binder, C.J. et al. Pneumococcal vaccination decreases atherosclerotic lesion formation: molecular mimicry between Streptococcus pneumoniae and oxidized LDL. Nat. Med. 9, 736–743 (2003).

    Article  CAS  PubMed  Google Scholar 

  39. Schmitz, G. & Grandl, M. Endolysosomal phospholipidosis and cytosolic lipid droplet storage and release in macrophages. Biochim. Biophys. Acta 1791, 524–539 (2009).

    Article  CAS  PubMed  Google Scholar 

  40. Rosenbaum, A.I. et al. Chemical screen to reduce sterol accumulation in Niemann-Pick C disease cells identifies novel lysosomal acid lipase inhibitors. Biochim. Biophys. Acta 1791, 1155–1165 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Heneka, M.T. et al. NLRP3 is activated in Alzheimer's disease and contributes to pathology in APP/PS1 mice. Nature 493, 674–678 (2013).

    Article  CAS  PubMed  Google Scholar 

  42. Friedrich, R.P. et al. Mechanism of amyloid plaque formation suggests an intracellular basis of Abeta pathogenicity. Proc. Natl. Acad. Sci. USA 107, 1942–1947 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Walsh, D.M., Tseng, B.P., Rydel, R.E., Podlisny, M.B. & Selkoe, D.J. The oligomerization of amyloid β-protein begins intracellularly in cells derived from human brain. Biochemistry 39, 10831–10839 (2000).

    Article  CAS  PubMed  Google Scholar 

  44. Miao, E.A. et al. Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat. Immunol. 11, 1136–1142 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Kamari, Y. et al. Reduced atherosclerosis and inflammatory cytokines in apolipoprotein-E-deficient mice lacking bone marrow-derived interleukin-1α. Biochem. Biophys. Res. Commun. 405, 197–203 (2011).

    Article  CAS  PubMed  Google Scholar 

  46. Kuchibhotla, S. et al. Absence of CD36 protects against atherosclerosis in ApoE knock-out mice with no additional protection provided by absence of scavenger receptor A I/II. Cardiovasc. Res. 78, 185–196 (2008).

    Article  CAS  PubMed  Google Scholar 

  47. Chen, C.J. et al. Identification of a key pathway required for the sterile inflammatory response triggered by dying cells. Nat. Med. 13, 851–856 (2007).

    Article  CAS  PubMed  Google Scholar 

  48. Rasmussen, L.T. & Seljelid, R. The modulatory effect of lipoproteins on the release of interleukin 1 by human peritoneal macrophages stimulated with β-1,3-D-polyglucose derivatives. Scand. J. Immunol. 29, 477–484 (1989).

    Article  CAS  PubMed  Google Scholar 

  49. Thomas, C.E., Jackson, R.L., Ohlweiler, D.F. & Ku, G. Multiple lipid oxidation products in low density lipoproteins induce interleukin-1β release from human blood mononuclear cells. J. Lipid Res. 35, 417–427 (1994).

    Article  CAS  PubMed  Google Scholar 

  50. Moyer, C.F., Sajuthi, D., Tulli, H. & Williams, J.K. Synthesis of IL-1α and IL-1β by arterial cells in atherosclerosis. Am. J. Pathol. 138, 951–960 (1991).

    CAS  PubMed  PubMed Central  Google Scholar 

  51. Guarda, G. & So, A. Regulation of inflammasome activity. Immunology 130, 329–336 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Mariathasan, S. et al. Cryopyrin activates the inflammasome in response to toxins and ATP. Nature 440, 228–232 (2006).

    Article  CAS  PubMed  Google Scholar 

  53. Boltz-Nitulescu, G. et al. Differentiation of rat bone marrow cells into macrophages under the influence of mouse L929 cell supernatant. J. Leuk. Biol. 41, 83–91 (1987).

    Article  CAS  Google Scholar 

  54. Stuart, L.M. et al. Response to Staphylococcus aureus requires CD36-mediated phagocytosis triggered by the COOH-terminal cytoplasmic domain. J. Cell Biol. 170, 477–485 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank F. Maxfield (Weill Cornell Medical College) for Lalistat. Supported by the US National Institutes of Health (R01HL117334 and R01AG032349 to K.J.M.; U24 AI082660 to L.M.S. and K.J.M.; R01 AI079198 to L.M.S.; 5R01HL093262-02 and 1R01HL112661-01 to E.L.; and AI083713 to K.A.F. and E.L.) and the American Heart Association (11POST7400075 to F.J.S.).

Author information

Authors and Affiliations

  1. Department of Medicine, The Leon H. Charney Division of Cardiology, Marc and Ruti Bell Program for Vascular Biology and Disease, New York University School of Medicine, New York, New York, USA

    Frederick J Sheedy, Katey J Rayner, Bhama Ramkhelawon, Hasini N Ediriweera & Kathryn J Moore

  2. Institute of Innate Immunity, University of Bonn, Bonn, Germany

    Alena Grebe & Eicke Latz

  3. Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Parisa Kalantari, Susan B Carpenter, Douglas T Golenbock, Eicke Latz & Katherine A Fitzgerald

  4. Department of Pediatrics, Laboratory of Developmental Immunology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Christine E Becker & Lynda M Stuart

  5. Isis Pharmaceuticals, Carlsbad, California, USA

    Adam E Mullick

  6. German Center for Neurodegenerative Diseases, Bonn, Germany

    Eicke Latz

Authors
  1. Frederick J Sheedy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alena Grebe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Katey J Rayner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Parisa Kalantari
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bhama Ramkhelawon
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Susan B Carpenter
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christine E Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hasini N Ediriweera
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Adam E Mullick
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Douglas T Golenbock
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Lynda M Stuart
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Eicke Latz
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Katherine A Fitzgerald
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Kathryn J Moore
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

F.J.S. designed, did and analyzed experiments; A.G. analyzed cholesterol crystals in atherosclerotic plaques; K.J.R. and H.N.E. assisted with mouse atherosclerosis experiments. P.K., B.R., S.B.C. and C.E.B. assisted with microscopy experiments; A.E.M. provided CD36-specific ASOs and contributed to experimental design; D.T.G., L.M.S., E.L. and K.A.F. contributed to study design, data analysis, and manuscript preparation; and K.J.M. conceived of the study, designed and analyzed experiments and, along with F.J.S., prepared the manuscript.

Corresponding author

Correspondence to Kathryn J Moore.

Ethics declarations

Competing interests

A.E.M. is an employee of Isis Pharmaceuticals, a biotechnology company that develops ASO therapies.

Supplementary information

Supplementary Figures and Tables

Supplementary Figures 1–6 and Supplementary Table 1 (PDF 9146 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sheedy, F., Grebe, A., Rayner, K. et al. CD36 coordinates NLRP3 inflammasome activation by facilitating intracellular nucleation of soluble ligands into particulate ligands in sterile inflammation. Nat Immunol 14, 812–820 (2013). https://doi.org/10.1038/ni.2639

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.2639

This article is cited by

Search

Advanced 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