Molecular Medicine

, Volume 18, Issue 2, pp 201–208 | Cite as

Interferon Regulatory Factor-1 Regulates the Autophagic Response in LPS-Stimulated Macrophages through Nitric Oxide

  • Lemeng Zhang
  • Jon S. Cardinal
  • Runalia Bahar
  • John Evankovich
  • Hai Huang
  • Gary Nace
  • Timothy R. Billiar
  • Matthew R. Rosengart
  • Pinhua Pan
  • Allan Tsung
Research Article


The pathogenesis of sepsis is complex and, unfortunately, poorly understood. The cellular process of autophagy is believed to play a protective role in sepsis; however, the mechanisms responsible for its regulation in this setting are ill defined. In the present study, interferon regulatory factor 1 (IRF-1) was found to regulate the autophagic response in lipopolysaccharide (LPS)-stimulated macrophages. In vivo, tissue macrophages obtained from LPS-stimulated IRF-1 knockout (KO) mice demonstrated increased autophagy and decreased apoptosis compared to those isolated from IRF-1 wild-type (WT) mice. In vitro, LPS-stimulated peritoneal macrophages obtained from IRF-1 KO mice experienced increased autophagy and decreased apoptosis. IRF-1 mediates the inhibition of autophagy by modulating the activation of the mammalian target of rapamycin (mTOR). LPS induced the activation of mTOR in WT peritoneal macrophages, but not in IRF-1 KO macrophages. In contrast, overexpression of IRF-1 alone increased the activation of mTOR and consequently decreased autophagic flux. Furthermore, the inhibitory effects of IRF-1 mTOR activity were mediated by nitric oxide (NO). Therefore, we propose a novel role for IRF-1 and NO in the regulation of macrophage autophagy during LPS stimulation in which IRF-1/NO inhibits autophagy through mTOR activation.


  1. 1.
    Anderson RN, Smith BL. (2005) Deaths: leading causes for 2002. Natl. Vital Stat. Rep. 53:1–89.PubMedGoogle Scholar
  2. 2.
    Chopra M, Sharma AC. (2007) Distinct cardiodynamic and molecular characteristics during early and late stages of sepsis-induced myocardial dysfunction. Life Sci. 81:306–16.CrossRefGoogle Scholar
  3. 3.
    Xiao H, Siddiqui J, Remick DG. (2006) Mechanisms of mortality in early and late sepsis. Infect. Immun. 74:5227–35.CrossRefGoogle Scholar
  4. 4.
    Rubinsztein DC, Gestwicki JE, Murphy LO, Klionsky DJ. (2007) Potential therapeutic applications of autophagy. Nat. Rev. Drug. Discov. 6:304–12.CrossRefGoogle Scholar
  5. 5.
    He C, Klionsky DJ. (2009) Regulation mechanisms and signaling pathways of autophagy. Annu. Rev. Genet. 43:67–93.CrossRefGoogle Scholar
  6. 6.
    Levine B, Kroemer G. (2008) Autophagy in the pathogenesis of disease. Cell. 132:27–42.CrossRefGoogle Scholar
  7. 7.
    Sanjuan MA, et al. (2007) Toll-like receptor signalling in macrophages links the autophagy pathway to phagocytosis. Nature. 450:1253–7.CrossRefGoogle Scholar
  8. 8.
    Levine B, Yuan J. (2005) Autophagy in cell death: an innocent convict? J. Clin. Invest. 115:2679–88.CrossRefGoogle Scholar
  9. 9.
    Virgin HW, Levine B. (2009) Autophagy genes in immunity. Nat. Immunol. 10:461–70.CrossRefGoogle Scholar
  10. 10.
    Waltz P, et al. (2010) Lipopolysaccaride induces autophagic signaling in macrophages via a TLR4, heme oxygenase-1 dependent pathway. Autophagy. 7:315–20.CrossRefGoogle Scholar
  11. 11.
    Saitoh T, et al. (2008) Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1beta production. Nature. 456:264–8.CrossRefGoogle Scholar
  12. 12.
    Hamacher-Brady A, Brady NR, Gottlieb RA. (2006) Enhancing macroautophagy protects against ischemia/reperfusion injury in cardiac myocytes. J. Biol. Chem. 281:29776–87.CrossRefGoogle Scholar
  13. 13.
    Xu Y, et al. (2007) Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity. 27:135–44.CrossRefGoogle Scholar
  14. 14.
    Carchman EH, Rao J, Loughran PA, Rosengart MR, Zuckerbraun BS. (2011) Heme oxygenase-1-mediated autophagy protects against hepatocyte cell death and hepatic injury from infection/sepsis in mice. Hepatology. 53:2053–62.CrossRefGoogle Scholar
  15. 15.
    Nakahira K, et al. (2010) Autophagy proteins regulate innate immune responses by inhibiting the release of mitochondrial DNA mediated by the NALP3 inflammasome. Nat. Immunol. 12:222–30.CrossRefGoogle Scholar
  16. 16.
    Gonzalez-Polo RA, et al. (2005) The apoptosis/autophagy paradox: autophagic vacuolization before apoptotic death. J. Cell Sci. 118:3091–102.CrossRefGoogle Scholar
  17. 17.
    Mizushima N, Levine B, Cuervo AM, Klionsky DJ. (2008) Autophagy fights disease through cellular self-digestion. Nature. 451:1069–75.CrossRefGoogle Scholar
  18. 18.
    Levine B, Deretic V. (2007) Unveiling the roles of autophagy in innate and adaptive immunity. Nat. Rev. Immunol. 7:767–77.CrossRefGoogle Scholar
  19. 19.
    Levine B. (2005) Eating oneself and uninvited guests: autophagy-related pathways in cellular defense. Cell. 120:159–62.PubMedGoogle Scholar
  20. 20.
    Kroger A, Koster M, Schroeder K, Hauser H, Mueller PP. (2002) Activities of IRF-1. J. Interferon Cytokine Res. 22:5–14.CrossRefGoogle Scholar
  21. 21.
    Kano A, Haruyama T, Akaike T, Watanabe Y. (1999) IRF-1 is an essential mediator in IFN-gamma-induced cell cycle arrest and apoptosis of primary cultured hepatocytes. Biochem. Biophys. Res. Commun. 257:672–7.CrossRefGoogle Scholar
  22. 22.
    Stang MT, et al. (2007) Interferon regulatory factor-1-induced apoptosis mediated by a ligand-independent fas-associated death domain pathway in breast cancer cells. Oncogene. 26:6420–30.CrossRefGoogle Scholar
  23. 23.
    Papageorgiou A, Dinney CP, McConkey DJ. (2007) Interferon-alpha induces TRAIL expression and cell death via an IRF-1-dependent mechanism in human bladder cancer cells. Cancer Biol. Ther. 6:872–9.CrossRefGoogle Scholar
  24. 24.
    Tamura T, et al. (1995) An IRF-1-dependent pathway of DNA damage-induced apoptosis in mitogen-activated T lymphocytes. Nature. 376:596–9.CrossRefGoogle Scholar
  25. 25.
    Senaldi G, et al. (1999) Protection against the mortality associated with disease models mediated by TNF and IFN-gamma in mice lacking IFN regulatory factor-1. J. Immunol. 163:6820–6.PubMedGoogle Scholar
  26. 26.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington, (DC): National Academies Press. 220 pp. Available from: Scholar
  27. 27.
    Nakamura K, Yamaji T, Crocker PR, Suzuki A, Hashimoto Y. (2002) Lymph node macrophages, but not spleen macrophages, express high levels of unmasked sialoadhesin: implication for the adhesive properties of macrophages in vivo. Glycobiology. 12:209–16.CrossRefGoogle Scholar
  28. 28.
    Zhang C, et al. (2006) Toll-like receptor 2 mediates alveolar macrophage response to Pneumocystis murina. Infect. Immun. 74:1857–64.CrossRefGoogle Scholar
  29. 29.
    Ayala A, Urbanich MA, Herdon CD, Chaudry IH. (1996) Is sepsis-induced apoptosis associated with macrophage dysfunction? J. Trauma 40:568–73; discussion 573–4.CrossRefGoogle Scholar
  30. 30.
    ten Hagen TL, van Vianen W, Bakker-Woudenberg IA. (1996) Isolation and characterization of murine Kupffer cells and splenic macrophages. J. Immunol. Methods 193:81–91.CrossRefGoogle Scholar
  31. 31.
    Lederer JA. (2005) Does nitric oxide control the counter-inflammatory response in endotoxic shock? Crit. Care Med. 33:896–8.CrossRefGoogle Scholar
  32. 32.
    Kamijo R, et al. (1994) Requirement for transcription factor IRF-1 in NO synthase induction in macrophages. Science. 263:1612–5.CrossRefGoogle Scholar
  33. 33.
    Gotoh T, Oyadomari S, Mori K, Mori M. (2002) Nitric oxide-induced apoptosis in RAW 264.7 macrophages is mediated by endoplasmic reticulum stress pathway involving ATF6 and CHOP. J. Biol. Chem. 277:12343–50.CrossRefGoogle Scholar
  34. 34.
    Hunter CJ, et al. (2009) Lactobacillus bulgaricus prevents intestinal epithelial cell injury caused by Enterobacter sakazakii-induced nitric oxide both in vitro and in the newborn rat model of necrotizing enterocolitis. Infect. Immun. 77:1031–43.CrossRefGoogle Scholar
  35. 35.
    Zamora R, et al. (2005) Intestinal and hepatic expression of BNIP3 in necrotizing enterocolitis: regulation by nitric oxide and peroxynitrite. Am. J. Physiol. Gastrointest. Liver Physiol. 289:G822–30.CrossRefGoogle Scholar
  36. 36.
    Sarkar S, et al. (2011) Complex inhibitory effects of nitric oxide on autophagy. Mol. Cell. 43:19–32.CrossRefGoogle Scholar
  37. 37.
    Angus DC, et al. (2001) Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care. Crit. Care Med. 29:1303–10.CrossRefGoogle Scholar
  38. 38.
    Ziegler EJ, et al. (1991) Treatment of gram-negative bacteremia and septic shock with HA-1A human monoclonal antibody against endotoxin. A randomized, double-blind, placebo-controlled trial. The HA-1A Sepsis Study Group. N. Engl. J. Med. 324:429–36.CrossRefGoogle Scholar
  39. 39.
    Fisher CJ Jr, et al. (1996) Treatment of septic shock with the tumor necrosis factor receptor: Fc fusion protein. The Soluble TNF Receptor Sepsis Study Group. N. Engl. J. Med. 334:1697–702.CrossRefGoogle Scholar
  40. 40.
    Fisher CJ Jr, et al. (1994) Initial evaluation of human recombinant interleukin-1 receptor antagonist in the treatment of sepsis syndrome: a randomized, open-label, placebo-controlled multicenter trial. Crit. Care Med. 22:12–21.CrossRefGoogle Scholar
  41. 41.
    Haldar SM, Stamler JS. (2011) S-nitrosylation at the interface of autophagy and disease. Mol. Cell. 43:1–3.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2012

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Lemeng Zhang
    • 1
    • 2
  • Jon S. Cardinal
    • 1
  • Runalia Bahar
    • 1
  • John Evankovich
    • 1
  • Hai Huang
    • 1
  • Gary Nace
    • 1
  • Timothy R. Billiar
    • 1
  • Matthew R. Rosengart
    • 1
  • Pinhua Pan
    • 1
    • 2
  • Allan Tsung
    • 1
    • 3
  1. 1.Department of SurgeryUniversity of PittsburghPittsburghUSA
  2. 2.Department of Pulmonology, Xiangya HospitalCentral South UniversityChangshaChina
  3. 3.Montefiore HospitalUPMC Liver Cancer CenterPittsburghUSA

Personalised recommendations