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CD14 regulates the dendritic cell life cycle after LPS exposure through NFAT activation

Nature volume 460, pages 264–268 (2009)Cite this article

Abstract

Toll-like receptors (TLRs) are the best characterized pattern recognition receptors1. Individual TLRs recruit diverse combinations of adaptor proteins, triggering signal transduction pathways and leading to the activation of various transcription factors, including nuclear factor κB, activation protein 1 and interferon regulatory factors2. Interleukin-2 is one of the molecules produced by mouse dendritic cells after stimulation by different pattern recognition receptor agonists3,4,5,6. By analogy with the events after T-cell receptor engagement leading to interleukin-2 production, it is therefore plausible that the stimulation of TLRs on dendritic cells may lead to activation of the Ca2+/calcineurin and NFAT (nuclear factor of activated T cells) pathway. Here we show that mouse dendritic cell stimulation with lipopolysaccharide (LPS) induces Src-family kinase and phospholipase Cγ2 activation, influx of extracellular Ca2+ and calcineurin-dependent nuclear NFAT translocation. The initiation of this pathway is independent of TLR4 engagement, and dependent exclusively on CD14. We also show that LPS-induced NFAT activation via CD14 is necessary to cause the apoptotic death of terminally differentiated dendritic cells, an event that is essential for maintaining self-tolerance and preventing autoimmunity7,8. Consequently, blocking this pathway in vivo causes prolonged dendritic cell survival and an increase in T-cell priming capability. Our findings reveal novel aspects of molecular signalling triggered by LPS in dendritic cells, and identify a new role for CD14: the regulation of the dendritic cell life cycle through NFAT activation. Given the involvement of CD14 in disease, including sepsis and chronic heart failure9,10, the discovery of signal transduction pathways activated exclusively via CD14 is an important step towards the development of potential treatments involving interference with CD14 functions.

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Figure 1: CD14-dependent Ca 2+ mobilization in dendritic cells following LPS treatment.
Figure 2: CD14-dependent activation of NFAT in BMDCs after LPS treatment.
Figure 3: Ca 2+ -NFAT-mediated regulation of dendritic cell death.
Figure 4: Ca 2+ mobilization and survival of macrophages after LPS treatment.

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Primary accessions

Gene Expression Omnibus

Data deposits

All microarray data are available from the Gene Expression Omnibus database (http://www.ncbi.nlm.nih.gov/geo) under accession code GSE15759.

References

  1. Akira, S. Toll-like receptors and innate immunity. Adv. Immunol. 78, 1–56 (2001)

    Article  CAS  Google Scholar 

  2. Kaisho, T. & Akira, S. Toll-like receptor function and signaling. J. Allergy Clin. Immunol. 117, 979–987 (2006)

    Article  CAS  Google Scholar 

  3. Granucci, F., Feau, S., Angeli, V., Trottein, F. & Ricciardi-Castagnoli, P. Early IL-2 production by mouse dendritic cells is the result of microbial-induced priming. J. Immunol. 170, 5075–5081 (2003)

    Article  CAS  Google Scholar 

  4. Guiducci, C., Valzasina, B., Dislich, H. & Colombo, M. P. CD40/CD40L interaction regulates CD4+CD25+ Treg homeostasis through dendritic cell-produced IL-2. Eur. J. Immunol. 35, 557–567 (2005)

    Article  CAS  Google Scholar 

  5. Rogers, N. C. et al. Syk-dependent cytokine induction by Dectin-1 reveals a novel pattern recognition pathway for C type lectins. Immunity 22, 507–517 (2005)

    Article  CAS  Google Scholar 

  6. Yamazaki, S. et al. Direct expansion of functional CD25+ CD4+regulatory T cells by antigen-processing dendritic cells. J. Exp. Med. 198, 235–247 (2003)

    Article  CAS  Google Scholar 

  7. Chen, M. et al. Dendritic cell apoptosis in the maintenance of immune tolerance. Science 311, 1160–1164 (2006)

    Article  ADS  CAS  Google Scholar 

  8. Stranges, P. B. et al. Elimination of antigen-presenting cells and autoreactive T cells by Fas contributes to prevention of autoimmunity. Immunity 26, 629–641 (2007)

    Article  CAS  Google Scholar 

  9. Van Amersfoort, E. S., Van Berkel, T. J. & Kuiper, J. Receptors, mediators, and mechanisms involved in bacterial sepsis and septic shock. Clin. Microbiol. Rev. 16, 379–414 (2003)

    Article  CAS  Google Scholar 

  10. Genth-Zotz, S. et al. The anti-CD14 antibody IC14 suppresses ex vivo endotoxin stimulated tumor necrosis factor-α in patients with chronic heart failure. Eur. J. Heart Fail. 8, 366–372 (2006)

    Article  CAS  Google Scholar 

  11. Beutler, B. et al. Genetic analysis of host resistance: Toll-like receptor signaling and immunity at large. Annu. Rev. Immunol. 24, 353–389 (2006)

    Article  CAS  Google Scholar 

  12. Winzler, C. et al. Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures. J. Exp. Med. 185, 317–328 (1997)

    Article  CAS  Google Scholar 

  13. Kawasaki, K., Gomi, K., Kawai, Y., Shiozaki, M. & Nishijima, M. Molecular basis for lipopolysaccharide mimetic action of Taxol and flavolipin. J. Endotoxin Res. 9, 301–307 (2003)

    Article  CAS  Google Scholar 

  14. Suzuki, K. G. et al. GPI-anchored receptor clusters transiently recruit Lyn and Gα for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1. J. Cell Biol. 177, 717–730 (2007)

    Article  CAS  Google Scholar 

  15. Carpenter, G. & Ji, Q. Phospholipase C-γ as a signal-transducing element. Exp. Cell Res. 253, 15–24 (1999)

    Article  CAS  Google Scholar 

  16. Pugin, J. et al. Cell activation mediated by glycosylphosphatidylinositol-anchored or transmembrane forms of CD14. Infect. Immun. 66, 1174–1180 (1998)

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Kinoshita, T., Fujita, M. & Maeda, Y. Biosynthesis, remodelling and functions of mammalian GPI-anchored proteins: recent progress. J. Biochem. 144, 287–294 (2008)

    Article  CAS  Google Scholar 

  18. Moore, K. J. et al. Divergent response to LPS and bacteria in CD14-deficient murine macrophages. J. Immunol. 165, 4272–4280 (2000)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  20. Jiang, Z. et al. CD14 is required for MyD88-independent LPS signaling. Nature Immunol. 6, 565–570 (2005)

    Article  CAS  Google Scholar 

  21. Kagan, J. C. et al. TRAM couples endocytosis of Toll-like receptor 4 to the induction of interferon-β. Nature Immunol. 9, 361–368 (2008)

    Article  CAS  Google Scholar 

  22. Shakhov, A. N., Collart, M. A., Vassalli, P., Nedospasov, S. A. & Jongeneel, C. V. κB-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor α gene in primary macrophages. J. Exp. Med. 171, 35–47 (1990)

    Article  CAS  Google Scholar 

  23. Dendorfer, U. Molecular biology of cytokines. Artif. Organs 20, 437–444 (1996)

    Article  CAS  Google Scholar 

  24. Aramburu, J. et al. Affinity-driven peptide selection of an NFAT inhibitor more selective than cyclosporin A. Science 285, 2129–2133 (1999)

    Article  CAS  Google Scholar 

  25. Schuh, K. et al. Retarded thymic involution and massive germinal center formation in NF-ATp-deficient mice. Eur. J. Immunol. 28, 2456–2466 (1998)

    Article  CAS  Google Scholar 

  26. Mahnke, K., Qian, Y., Knop, J. & Enk, A. H. Induction of CD4+/CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 101, 4862–4869 (2003)

    Article  CAS  Google Scholar 

  27. Dobrovolskaia, M. A. & Vogel, S. N. Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect. 4, 903–914 (2002)

    Article  CAS  Google Scholar 

  28. Medzhitov, R. Origin and physiological roles of inflammation. Nature 454, 428–435 (2008)

    Article  ADS  CAS  Google Scholar 

  29. Granucci, F. et al. Inducible IL-2 production by dendritic cells revealed by global gene expression analysis. Nature Immunol. 2, 882–888 (2001)

    Article  CAS  Google Scholar 

  30. Schreiber, E., Matthias, P., Muller, M. M. & Schaffner, W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 17, 6419 (1989)

    Article  CAS  Google Scholar 

  31. Granucci, F. et al. The scavenger receptor MARCO mediates cytoskeleton rearrangements in dendritic cells and microglia. Blood 102, 2940–2947 (2003)

    Article  CAS  Google Scholar 

  32. Irizarry, R. A. et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249–264 (2003)

    Article  Google Scholar 

  33. Gentleman, R. C. et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004)

    Article  Google Scholar 

Download references

Acknowledgements

We thank S. Akira, C. Kirschning and K. Miyake for mutant mice; M. Colonna for mutant mice and advice on PLC-γ2 activation; E. Serfling for mutant mice and advice on NFAT activation; A. Rao for the VIVIT peptide. We also thank Genopolis–Consorzio di Genomica Funzionale for microarray hybridization and data analysis and K. Mahnke for the anti-DEC205–OVA conjugate. This work was supported by grants from the CARIPLO Foundation, the European Commission 6th Framework Program (MUGEN and DC-THERA contracts), the European Commission 7th Framework Program (TOLERAGE and ENCITE contracts), the Associazione Italiana per la Ricerca sul Cancro (AIRC) and the and the Italian Ministry of Education and Research (COFIN).

Author Contributions F.G. conceived and oversaw the project and wrote the paper; I.Z. conceived the research and conducted most of the experiments with R.O.; M.C., M.C., M.R., G.C., B.C. and A.Z. helped with calcium experiments; G.C. and F.M. helped with experiments on D1 cells; M.F. helped with quantitative real-time PCR; A.E.R. performed the EMSA experiments; P.R.-C. provided advice.

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Author notes

  1. Paola Ricciardi-Castagnoli

    Present address: Present address: Singapore Immunology Network (SIgN), Biomedical Sciences Institutes, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, IMMUNOS, 138648, Singapore.,

Authors and Affiliations

  1. Department of Biotechnology and Bioscience, University of Milano-Bicocca, Piazza della Scienza 2, 20126 Milan, Italy,

    Ivan Zanoni, Renato Ostuni, Giusy Capuano, Antonella Ellena Ronchi, Marcella Rocchetti, Francesca Mingozzi, Maria Foti, Barbara Costa, Antonio Zaza, Paola Ricciardi-Castagnoli & Francesca Granucci

  2. Department of Physics, University of Milano-Bicocca, Piazza della Scienza 3, 20126 Milan, Italy,

    Maddalena Collini, Michele Caccia & Giuseppe Chirico

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  1. Ivan Zanoni
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Corresponding author

Correspondence to Francesca Granucci.

Supplementary information

Supplementary Information

This file contains Supplementary Figures S1-S14, Supplementary Tables 1-2 and Supplementary References. (PDF 3307 kb)

Supplementary Movie 1

Movie BMDC_ATP shows Ca2+ mobilization in BMDCs after steady-state superfusion with ATP. (MOV 2391 kb)

Supplementary Movie 2

Movie BMDC_LPS shows Ca2+ mobilization in BMDCs after steady-state superfusion with LPS. (MOV 1823 kb)

Supplementary Movie 3

Movie DI_ATP shows Ca2+ mobilization in D1 cells after steady-state superfusion with ATP. (MOV 3312 kb)

Supplementary Movie 4

Movie DI_LPS shows Ca2+ mobilization in D1 cells after steady-state superfusion with LPS. (MOV 2042 kb)

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Zanoni, I., Ostuni, R., Capuano, G. et al. CD14 regulates the dendritic cell life cycle after LPS exposure through NFAT activation. Nature 460, 264–268 (2009). https://doi.org/10.1038/nature08118

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