Article | Published:

Glycolipid iGb3 feedback amplifies innate immune responses via CD1d reverse signaling

Cell Researchvolume 29pages4253 (2019) | Download Citation

Abstract

The cross-talk between cellular lipid metabolism and the innate immune responses remains obscure. In addition to presenting lipid antigens to Natural Killer T-cells (NKT cells), the Cluster of Differentiation 1D Glycoprotein (CD1d) might mediate reverse signaling in antigen-presenting cells (APCs). Here we found CD1d deficiency attenuated Toll-like receptor (TLR)-triggered inflammatory innate responses in macrophages and dendritic cells, protecting mice from endotoxin shock. TLR activation in macrophages induced metabolic changes of glycosphingolipids (GSLs), among which glycolipid isoglobotrihexosylceramide (iGb3) was rapidly produced. The endogenously generated iGb3 bound CD1d in endosomal compartments and then synergized with the initially activated TLR signal to induce Tyr332 phosphorylation of CD1d intracellular domain. This led to the recruitment and activation of proline-rich tyrosine kinase 2 (Pyk2). Pyk2 interacted with IκB kinase β (IKKβ) and TANK-binding kinase 1 (TBK1), and enhanced tyrosine phosphorylation of Tyr188/199 of IKKβ and Tyr179 of TBK1 and thus, their activation to promote full activation of TLR signaling. Thus, intracellular CD1d reverse signaling, triggered by endogenous iGb3, amplifies inflammatory innate responses in APCs. Our findings identify a non-canonical function of CD1d reverse signaling activated by lipid metabolite in the innate immune response.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Buck, M. D., Sowell, R. T., Kaech, S. M. & Pearce, E. L. Metabolic instruction of immunity. Cell 169, 570–586 (2017).

  2. 2.

    Ganeshan, K. & Chawla, A. Metabolic regulation of immune responses. Annu. Rev. Immunol. 32, 609–634 (2014).

  3. 3.

    Ghesquiere, B., Wong, B. W., Kuchnio, A. & Carmeliet, P. Metabolism of stromal and immune cells in health and disease. Nature 511, 167–176 (2014).

  4. 4.

    Brestoff, J. R. & Artis, D. Immune regulation of metabolic homeostasis in health and disease. Cell 161, 146–160 (2015).

  5. 5.

    McNelis, J. C. & Olefsky, J. M. Macrophages, immunity, and metabolic disease. Immunity 41, 36–48 (2014).

  6. 6.

    O’Neill, L. A. & Pearce, E. J. Immunometabolism governs dendritic cell and macrophage function. J. Exp. Med. 213, 15–23 (2016).

  7. 7.

    Phan, A. T., Goldrath, A. W. & Glass, C. K. Metabolic and epigenetic coordination of T cell and macrophage immunity. Immunity 46, 714–729 (2017).

  8. 8.

    Everts, B. et al. TLR-driven early glycolytic reprogramming via the kinases TBK1-IKKε supports the anabolic demands of dendritic cell activation. Nat. Immunol. 15, 323–332 (2014).

  9. 9.

    Pearce, E. J. & Everts, B. Dendritic cell metabolism. Nat. Rev. Immunol. 15, 18–29 (2015).

  10. 10.

    Andreyev, A. Y. et al. Subcellular organelle lipidomics in TLR-4-activated macrophages. J. Lip. Res. 51, 2785–2797 (2010).

  11. 11.

    Dennis, E. A. et al. A mouse macrophage lipidome. J. Biol. Chem. 285, 39976–39985 (2010).

  12. 12.

    Blanc, M. et al. The transcription factor STAT-1 couples macrophage synthesis of 25-hydroxycholesterol to the interferon antiviral response. Immunity 38, 106–118 (2013).

  13. 13.

    Coulombe, F. et al. Targeted prostaglandin E2 inhibition enhances antiviral immunity through induction of type I interferon and apoptosis in macrophages. Immunity 40, 554–568 (2014).

  14. 14.

    McEwen-Smith, R. M., Salio, M. & Cerundolo, V. CD1d-dependent endogenous and exogenous lipid antigen presentation. Curr. Opin. Immunol. 34, 116–125 (2015).

  15. 15.

    Zhou, D. et al. Lysosomal glycosphingolipid recognition by NKT cells. Science 306, 1786–1789 (2004).

  16. 16.

    Li, Y. et al. Immunologic glycosphingolipidomics and NKT cell development in mouse thymus. J. Proteome Res. 8, 2740–2751 (2009).

  17. 17.

    Paget, C. et al. Activation of invariant NKT cells by toll-like receptor 9-stimulated dendritic cells requires type I interferon and charged glycosphingolipids. Immunity 27, 597–609 (2007).

  18. 18.

    Salio, M., Silk, J. D., Jones, E. Y. & Cerundolo, V. Biology of CD1- and MR1-restricted T cells. Annu. Rev. Immunol. 32, 323–366 (2014).

  19. 19.

    Rossjohn, J., Pellicci, D. G., Patel, O., Gapin, L. & Godfrey, D. I. Recognition of CD1d-restricted antigens by natural killer T cells. Nat. Rev. Immunol. 12, 845–857 (2012).

  20. 20.

    Moody, D. B. TLR gateways to CD1 function. Nat. Immunol. 7, 811–817 (2006).

  21. 21.

    Kasmar, A., Van, Rhijn., I. & Moody, D. B. The evolved functions of CD1 during infection. Curr. Opin. Immunol. 21, 397–403 (2009).

  22. 22.

    Colgan, S. P., Hershberg, R. M., Furuta, G. T. & Blumberg, R. S. Ligation of intestinal epithelial CD1d induces bioactive IL-10: critical role of the cytoplasmic tail in autocrine signaling. Proc. Natl Acad. Sci. USA 96, 13938–13943 (1999).

  23. 23.

    Olszak, T. et al. Protective mucosal immunity mediated by epithelial CD1d and IL-10. Nature 509, 497–502 (2014).

  24. 24.

    Yue, S. C., Shaulov, A., Wang, R., Balk, S. P. & Exley, M. A. CD1d ligation on human monocytes directly signals rapid NF-κB activation and production of bioactive IL-12. Proc. Natl Acad. Sci. USA 102, 11811–11816 (2005).

  25. 25.

    Yue, S. C. et al. Direct CD1d-mediated stimulation of APC IL-12 production and protective immune response to virus infection in vivo. J. Immunol. 184, 268–276 (2010).

  26. 26.

    Brubaker, S. W., Bonham, K. S., Zanoni, I. & Kagan, J. C. Innate immune pattern recognition: a cell biological perspective. Annu. Rev. Immunol. 33, 257–290 (2015).

  27. 27.

    Cao, X. Self-regulation and cross-regulation of pattern-recognition receptor signalling in health and disease. Nat. Rev. Immunol. 16, 35–50 (2016).

  28. 28.

    O’Neill, L. A., Golenbock, D. & Bowie, A. G. The history of Toll-like receptors- redefining innate immunity. Nat. Rev. Immunol. 13, 453–460 (2013).

  29. 29.

    Liu, X. et al. Intracellular MHC class II molecules promote TLR-triggered innate immune responses by maintaining activation of the kinase Btk. Nat. Immunol. 12, 416–424 (2011).

  30. 30.

    Xu, S. et al. Constitutive MHC class I molecules negatively regulate TLR-triggered inflammatory responses via the Fps-SHP-2 pathway. Nat. Immunol. 13, 551–559 (2012).

  31. 31.

    Haziot, A. et al. Resistance to endotoxin shock and reduced dissemination of gram-negative bacteria in CD14-deficient mice. Immunity 4, 407–414 (1996).

  32. 32.

    Speak, A. O. et al. Implications for invariant natural killer T cell ligands due to the restricted presence of isoglobotrihexosylceramide in mammals. Proc. Natl Acad. Sci. USA 104, 5971–5976 (2007).

  33. 33.

    Mattner, J. et al. Exogenous and endogenous glycolipid antigens activate NKT cells during microbial infections. Nature 434, 525–529 (2005).

  34. 34.

    Xia, C. et al. Synthesis and biological evaluation of alpha-galactosylceramide (KRN7000) and isoglobotrihexosylceramide (iGb3). Bioorg. Med. Chem. Lett. 16, 2195–2199 (2006).

  35. 35.

    Kain, L. et al. The identification of the endogenous ligands of natural killer T cells reveals the presence of mammalian alpha-linked glycosylceramides. Immunity 41, 543–554 (2014).

  36. 36.

    Monzon-Casanova, E. et al. CD1d expression in paneth cells and rat exocrine pancreas revealed by novel monoclonal antibodies which differentially affect NKT cell activation. PLoS ONE 5, e13089 (2010).

  37. 37.

    Rodionov, D. G., Nordeng, T. W., Pedersen, K., Balk, S. P. & Bakke, O. A critical tyrosine residue in the cytoplasmic tail is important for CD1d internalization but not for its basolateral sorting in MDCK cells. J. Immunol. 162, 1488–1495 (1999).

  38. 38.

    Kamen, L. A., Schlessinger, J. & Lowell, C. A. Pyk2 is required for neutrophil degranulation and host defense responses to bacterial infection. J. Immunol. 186, 1656–1665 (2011).

  39. 39.

    Okigaki, M. et al. Pyk2 regulates multiple signaling events crucial for macrophage morphology and migration. Proc. Natl Acad. Sci. USA 100, 10740–10745 (2003).

  40. 40.

    Duan, Y., Learoyd, J., Meliton, A. Y., Leff, A. R. & Zhu, X. Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury. Resp. Res. 13, 4 (2012).

  41. 41.

    Zhang, P. et al. Protein tyrosine phosphatase with proline-glutamine-serine-threonine-rich motifs negatively regulates TLR-triggered innate responses by selectively inhibiting IκB kinase β/NF-κB activation. J. Immunol. 190, 1685–1694 (2013).

  42. 42.

    Larabi, A. et al. Crystal structure and mechanism of activation of TANK-binding kinase 1. Cell Rep. 3, 734–746 (2013).

  43. 43.

    Li, X. et al. The tyrosine kinase Src promotes phosphorylation of the kinase TBK1 to facilitate type I interferon production after viral infection. Sci. Signal. 10, eaae0435 (2017).

  44. 44.

    Chaudhry, M. S. & Karadimitris, A. Role and regulation of CD1d in normal and pathological B cells. J. Immunol. 193, 4761–4768 (2014).

  45. 45.

    Kojo, S., Tsutsumi., A., Goto, D. & Sumida, T. Low expression levels of soluble CD1d gene in patients with rheumatoid arthritis. J. Rheum. 30, 2524–2528 (2003).

  46. 46.

    Maceyka, M. & Spiegel, S. Sphingolipid metabolites in inflammatory disease. Nature 510, 58–67 (2014).

  47. 47.

    Perry, R. J., Samuel, V. T., Petersen, K. F. & Shulman, G. I. The role of hepatic lipids in hepatic insulin resistance and type 2 diabetes. Nature 510, 84–91 (2014).

  48. 48.

    Buckley, C. D., Gilroy, D. W. & Serhan, C. N. Proresolving lipid mediators and mechanisms in the resolution of acute inflammation. Immunity 40, 315–327 (2014).

  49. 49.

    Kelly, B. & O’Neill, L. A. Metabolic reprogramming in macrophages and dendritic cells in innate immunity. Cell Res. 25, 771–784 (2015).

  50. 50.

    Darmoise, A. et al. Lysosomal alpha-galactosidase controls the generation of self lipid antigens for natural killer T cells. Immunity 33, 216–228 (2010).

  51. 51.

    Li, Y., Zhou, D., Xia, C., Wang, P. G. & Levery, S. B. Sensitive quantitation of isoglobotriaosylceramide in the presence of isobaric components using electrospray ionization-ion trap mass spectrometry. Glycobiology 18, 166–176 (2008).

  52. 52.

    Li, Y. et al. Sensitive detection of isoglobo and globo series tetraglycosylceramides in human thymus by ion trap mass spectrometry. Glycobiology 18, 158–165 (2008).

  53. 53.

    Zajonc, D. M., Savage, P. B., Bendelac, A., Wilson, I. A. & Teyton, L. Crystal structures of mouse CD1d-iGb3 complex and its cognate Valpha14 T cell receptor suggest a model for dual recognition of foreign and self glycolipids. J. Mol. Biol. 377, 1104–1116 (2008).

  54. 54.

    Kang, S. J. & Cresswell, P. Regulation of intracellular trafficking of human CD1d by association with MHC class II molecules. EMBO J. 21, 1650–1660 (2002).

  55. 55.

    Zhang, Q. et al. Tet2 is required to resolve inflammation by recruiting Hdac2 to specifically repress IL-6. Nature 525, 389–393 (2015).

  56. 56.

    Wang, P. et al. The STAT3-binding long noncoding RNA lnc-DC controls human dendritic cell differentiation. Science 344, 310–313 (2014).

Download references

Acknowledgements

We thank Dr. H. Shen (University of Pennsylvania) for Listeria monocytogenes; and X. Sun and M. Jin for technical assistance. This work was supported by the National Key Basic Research Program of China (2015CB964403 to X.C.), the National Natural Science Foundation of China (81788101 to X.C., 31570871 to X.L., 31770970 to X.L., 81600182 to P.Z., 81571543 to Y.L.), CAMS Innovation Fund for Medical Sciences (2016-12M-1-003 to X.C.), and “Shuguang Program” of Shanghai Education Development Foundation and Shanghai Municipal Education Commission (18SG33 to X.L.).

Author information

Author notes

  1. These authors contributed equally: Xingguang Liu, Peng Zhang, Yunkai Zhang

Affiliations

  1. National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai, 200433, China

    • Xingguang Liu
    • , Peng Zhang
    • , Yunkai Zhang
    • , Sheng Xu
    • , Yingke Li
    • , Qingqing Zhou
    • , Xiang Chen
    • , Nan Li
    •  & Xuetao Cao
  2. Department of Haematology, General Hospital of Southern Theater Command, Guangzhou, 510000, China

    • Peng Zhang
  3. Laboratory of Cellular and Molecular Tumor Immunology, Institutes of Biology and Medical Sciences, Jiangsu Laboratory of Infection Immunity, Soochow University, Suzhou, 215123, China

    • Zheng Wang
    •  & Yunsen Li
  4. Institute of Immunology, Zhejiang University School of Medicine, Hangzhou, 310058, China

    • Wanwan Huai
    • , Xi Chen
    •  & Xuetao Cao
  5. College of Pharmacy, Nankai University, Tianjin, 30071, China

    • Peng Wang
  6. College of Life Science, Nankai University, Tianjin, 30071, China

    • Xuetao Cao
  7. Department of Immunology & Center for Immunotherapy, Institute of Basic Medical Sciences, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100005, China

    • Xuetao Cao

Authors

  1. Search for Xingguang Liu in:

  2. Search for Peng Zhang in:

  3. Search for Yunkai Zhang in:

  4. Search for Zheng Wang in:

  5. Search for Sheng Xu in:

  6. Search for Yingke Li in:

  7. Search for Wanwan Huai in:

  8. Search for Qingqing Zhou in:

  9. Search for Xiang Chen in:

  10. Search for Xi Chen in:

  11. Search for Nan Li in:

  12. Search for Peng Wang in:

  13. Search for Yunsen Li in:

  14. Search for Xuetao Cao in:

Contributions

X.C. and X.L. conceived this project and supervised the experiments. X.C., X.L., P.Z. and Y.Z. wrote the paper. X.L., P.Z., Y.Z., Z.W., S.X., Y.L., W.H., Q.Z., X.C., X.C., N.L. P.W. and Y.L. performed the experiments and analyzed the data.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Xingguang Liu or Xuetao Cao.

Electronic supplementary material

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/s41422-018-0122-7