Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1β production (original) (raw)

Nature volume 456, pages 264–268 (2008)Cite this article

Abstract

Systems for protein degradation are essential for tight control of the inflammatory immune response1,2. Autophagy, a bulk degradation system that delivers cytoplasmic constituents into autolysosomes, controls degradation of long-lived proteins, insoluble protein aggregates and invading microbes, and is suggested to be involved in the regulation of inflammation3,4,5. However, the mechanism underlying the regulation of inflammatory response by autophagy is poorly understood. Here we show that Atg16L1 (autophagy-related 16-like 1), which is implicated in Crohn's disease6,7, regulates endotoxin-induced inflammasome activation in mice. Atg16L1-deficiency disrupts the recruitment of the Atg12-Atg5 conjugate to the isolation membrane, resulting in a loss of microtubule-associated protein 1 light chain 3 (LC3) conjugation to phosphatidylethanolamine. Consequently, both autophagosome formation and degradation of long-lived proteins are severely impaired in Atg16L1-deficient cells. Following stimulation with lipopolysaccharide, a ligand for Toll-like receptor 4 (refs 8, 9), Atg16L1-deficient macrophages produce high amounts of the inflammatory cytokines IL-1β and IL-18. In lipopolysaccharide-stimulated macrophages, Atg16L1-deficiency causes Toll/IL-1 receptor domain-containing adaptor inducing IFN-β (TRIF)-dependent activation of caspase-1, leading to increased production of IL-1β. Mice lacking Atg16L1 in haematopoietic cells are highly susceptible to dextran sulphate sodium-induced acute colitis, which is alleviated by injection of anti-IL-1β and IL-18 antibodies, indicating the importance of Atg16L1 in the suppression of intestinal inflammation. These results demonstrate that Atg16L1 is an essential component of the autophagic machinery responsible for control of the endotoxin-induced inflammatory immune response.

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

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

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

Additional access options:

Similar content being viewed by others

References

  1. Liu, Y. C., Penninger, J. & Karin, M. Immunity by ubiquitylation: A reversible process of modification. Nature Rev. Immunol. 5, 941–952 (2005)
    Article CAS Google Scholar
  2. Wang, Y. et al. Lysosome-associated small Rab GTPase Rab7b negatively regulates TLR4 signaling in macrophages by promoting lysosomal degradation of TLR4. Blood 110, 962–971 (2007)
    Article CAS Google Scholar
  3. Ohsumi, Y. Molecular dissection of autophagy: Two ubiquitin-like systems. Nature Rev. Mol. Cell Biol. 2, 211–216 (2001)
    Article CAS Google Scholar
  4. Mizushima, N., Levine, B., Cuervo, A. M. & Klionsky, D. J. Autophagy fights disease through cellular self-digestion. Nature 451, 1069–1075 (2008)
    Article ADS CAS Google Scholar
  5. Levine, B. & Deretic, V. Unveiling the roles of autophagy in innate and adaptive immunity. Nature Rev. Immunol. 7, 767–777 (2007)
    Article CAS Google Scholar
  6. Hampe, J. et al. A genome-wide association scan of nonsynonymous SNPs identifies a susceptibility variant for Crohn disease in ATG16L1. Nature Genet. 39, 207–211 (2007)
    Article CAS Google Scholar
  7. Rioux, J. D. et al. Genome-wide association study identifies new susceptibility loci for Crohn disease and implicates autophagy in disease pathogenesis. Nature Genet. 39, 596–604 (2007)
    Article CAS Google Scholar
  8. Akira, S., Uematsu, S. & Takeuchi, O. Pathogen recognition and innate immunity. Cell 124, 783–801 (2006)
    Article CAS Google Scholar
  9. Yamamoto, M. et al. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301, 640–643 (2003)
    Article ADS CAS Google Scholar
  10. Mizushima, N. et al. Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate. J. Cell Sci. 116, 1679–1688 (2003)
    Article CAS Google Scholar
  11. Fujita, N., Itoh, T., Fukuda, M., Noda, T. & Yoshimori, T. The Atg16L complex specifies the site of LC3 lipidation for membrane biogenesis in autophagy. Mol. Biol. Cell 19, 2092–2100 (2008)
    Article CAS Google Scholar
  12. Kuma, A. et al. The role of autophagy during the early neonatal starvation period. Nature 432, 1032–1036 (2004)
    Article ADS CAS Google Scholar
  13. Komatsu, M. et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J. Cell Biol. 169, 425–434 (2005)
    Article CAS Google Scholar
  14. Lee, H. K., Lund, J. M., Ramanathan, B., Mizushima, N. & Iwasaki, A. Autophagy-dependent viral recognition by plasmacytoid dendritic cells. Science 315, 1398–1401 (2007)
    Article ADS CAS Google Scholar
  15. Sanjuan, M. A. et al. Toll-like receptor signalling in macrophages links the autophagy pathway to phagocytosis. Nature 450, 1253–1257 (2007)
    Article ADS CAS Google Scholar
  16. Kanneganti, T. D., Lamkanfi, M. & Núñez, G. Intracellular NOD-like receptors in host defense and disease. Immunity 27, 549–559 (2007)
    Article CAS Google Scholar
  17. Pétrilli, V., Dostert, C., Muruve, D. A. & Tschopp, J. The inflammasome: A danger sensing complex triggering innate immunity. Curr. Opin. Immunol. 19, 615–622 (2007)
    Article Google Scholar
  18. Hsu, L. C. et al. The protein kinase PKR is required for macrophage apoptosis after activation of Toll-like receptor 4. Nature 428, 341–345 (2004)
    Article ADS CAS Google Scholar
  19. Greten, F. R. et al. NF-κB is a negative regulator of IL-1β secretion as revealed by genetic and pharmacological inhibition of IKKβ. Cell 130, 918–931 (2007)
    Article CAS Google Scholar
  20. Dostert, C. et al. Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 320, 674–677 (2008)
    Article ADS CAS Google Scholar
  21. Hewinson, J., Moore, S. F., Glover, C., Watts, A. G. & MacKenzie, A. B. A key role for redox signaling in rapid P2X7 receptor-induced IL-1beta processing in human monocytes. J. Immunol. 180, 8410–8420 (2008)
    Article CAS Google Scholar
  22. Xu, Y. et al. Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity 27, 135–144 (2007)
    Article CAS Google Scholar
  23. Delgado, M. A., Elmaoued, R. A., Davis, A. S., Kyei, G. & Deretic, V. Toll-like receptors control autophagy. EMBO J. 27, 1110–1121 (2008)
    Article CAS Google Scholar
  24. Hara, T. et al. Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature 441, 885–889 (2006)
    Article ADS CAS Google Scholar
  25. Komatsu, M. et al. Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-deficient mice. Cell 131, 1149–1163 (2007)
    Article CAS Google Scholar
  26. Paludan, C. et al. Endogenous MHC class II processing of a viral nuclear antigen after autophagy. Science 307, 593–596 (2005)
    Article ADS CAS Google Scholar
  27. Maeda, S. et al. Nod2 mutation in Crohn's disease potentiates NF-κB activity and IL-1β processing. Science 307, 737–738 (2005)
    ADS Google Scholar
  28. Ishikura, T. et al. Interleukin-18 overproduction exacerbates the development of colitis with markedly infiltrated macrophages in interleukin-18 transgenic mice. J. Gastroenterol. Hepatol. 18, 960–969 (2003)
    Article CAS Google Scholar
  29. Izcue, A., Coombes, J. L. & Powrie, F. Regulatory T cells suppress systemic and mucosal immune activation to control intestinal inflammation. Immunol. Rev. 212, 256–271 (2006)
    Article CAS Google Scholar
  30. Nakagawa, I. et al. Autophagy defends cells against invading group A Streptococcus. Science 306, 1037–1040 (2004)
    Article ADS CAS Google Scholar
  31. Fujita, N. et al. An Atg4B mutant hampers the lipidation of LC3 paralogues and causes defects in autophagosome closure. Mol. Biol. Cell advance online publication, doi: 10.1091/mbc.E08-03-0312 (3 September 2008)

Download references

Acknowledgements

We are grateful to T. Kitamura, S. Yamaoka and N. Mizushima for providing materials. We thank K. J. Ishii, M. Yamamoto and members of the Laboratory of Host Defense for discussions; Y. Fujiwara, M. Shiokawa, R. Nakayama and N. Kitagaki for technical assistance; and M. Hashimoto and E. Kamada for secretarial assistance. This work was in part supported by grants from NIH (AI070167) and the Ministry of Health, Labour and Welfare of Japan, and by Grant-in-Aid for Specially Promoted Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author Contributions T.S. generated the Atg16L1-deficient mice and performed the immunological experiments. N.F. performed the cell biology experiments. N.Y. generated the retroviral vector. M.K. and K.T. generated the Atg7-deficient mice. T.T. performed histological analysis of mice. M.H.J., S.U., B.-G.Y., T.S., H.O., T.N., T.K. and O.T. helped with experiments. T.Y. designed the cell biology research. S.A. supervised the overall research project.

Author information

Author notes

  1. Tatsuya Saitoh and Naonobu Fujita: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Host Defense,,
    Tatsuya Saitoh, Satoshi Uematsu, Bo-Gie Yang, Takashi Satoh, Taro Kawai, Osamu Takeuchi & Shizuo Akira
  2. Laboratory of Gastrointestinal Immunology, WPI Immunology Frontier Research Center, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan ,
    Myoung Ho Jang
  3. Department of Host Defense,,
    Tatsuya Saitoh, Satoshi Uematsu, Bo-Gie Yang, Takashi Satoh, Taro Kawai, Osamu Takeuchi & Shizuo Akira
  4. Department of Cellular Regulation, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan,
    Naonobu Fujita, Hiroko Omori, Takeshi Noda & Tamotsu Yoshimori
  5. AIDS Research Center, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan ,
    Naoki Yamamoto
  6. Laboratory of Frontier Science, Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, Tokyo 113-8613, Japan ,
    Masaaki Komatsu & Keiji Tanaka
  7. Department of Biochemistry, Juntendo University School of Medicine, 2-1-1 Hongo Bunkyo-ku, Tokyo 113-8421, Japan,
    Masaaki Komatsu
  8. PRESTO, Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan ,
    Masaaki Komatsu
  9. Department of Pathology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan,
    Tohru Tsujimura
  10. CREST, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan ,
    Tamotsu Yoshimori

Authors

  1. Tatsuya Saitoh
    You can also search for this author inPubMed Google Scholar
  2. Naonobu Fujita
    You can also search for this author inPubMed Google Scholar
  3. Myoung Ho Jang
    You can also search for this author inPubMed Google Scholar
  4. Satoshi Uematsu
    You can also search for this author inPubMed Google Scholar
  5. Bo-Gie Yang
    You can also search for this author inPubMed Google Scholar
  6. Takashi Satoh
    You can also search for this author inPubMed Google Scholar
  7. Hiroko Omori
    You can also search for this author inPubMed Google Scholar
  8. Takeshi Noda
    You can also search for this author inPubMed Google Scholar
  9. Naoki Yamamoto
    You can also search for this author inPubMed Google Scholar
  10. Masaaki Komatsu
    You can also search for this author inPubMed Google Scholar
  11. Keiji Tanaka
    You can also search for this author inPubMed Google Scholar
  12. Taro Kawai
    You can also search for this author inPubMed Google Scholar
  13. Tohru Tsujimura
    You can also search for this author inPubMed Google Scholar
  14. Osamu Takeuchi
    You can also search for this author inPubMed Google Scholar
  15. Tamotsu Yoshimori
    You can also search for this author inPubMed Google Scholar
  16. Shizuo Akira
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toShizuo Akira.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

Saitoh, T., Fujita, N., Jang, M. et al. Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1β production.Nature 456, 264–268 (2008). https://doi.org/10.1038/nature07383

Download citation

This article is cited by

Editorial Summary

Inflammatory bowel disease

Crohn's disease, a chronic inflammation of the gut, has been linked to over thirty gene loci. Two papers in this issue focus a recent addition to that list, ATG16L1 (Atg16-like 1). Atg16 protein itself was first identified in yeast as an essential gene for the process of autophagy, a system that clears away unwanted cellular components and is involved in the pathogenesis of microbial infection, neurodegeneration and tumorigenesis. Cadwell et al. report a unique role for Atg16L1 in Paneth cells, a type of epithelial cell that secretes granules containing antimicrobial peptides into the intestines. Saitoh et al. show that ATG16L1 plays a role in the inflammatory response in isolated macrophages and in the mouse intestine, as an essential component of the autophagic machinery. This work implicates Atg16L1 in the control of inflammatory immune response and the maintenance of intestinal barrier, both of which are important for the prevention of inflammatory bowel disease.