Advertisement

Molecular Medicine

, Volume 18, Issue 3, pp 477–485 | Cite as

Inhibition of High-Mobility Group Box 1 Protein (HMGB1) Enhances Bacterial Clearance and Protects against Pseudomonas Aeruginosa Pneumonia in Cystic Fibrosis

  • Maria Entezari
  • Daniel J Weiss
  • Ravikumar Sitapara
  • Laurie Whittaker
  • Matthew J Wargo
  • JianHua Li
  • Haichao Wang
  • Huan Yang
  • Lokesh Sharma
  • Binh D Phan
  • Mohammad Javdan
  • Sangeeta S Chavan
  • Edmund J Miller
  • Kevin J Tracey
  • Lin L Mantell
Research Article

Abstract

Pulmonary infection with Pseudomonas aeruginosa and neutrophilic lung inflammation significantly contribute to morbidity and mortality in cystic fibrosis (CF). High-mobility group box 1 protein (HMGB1), a ubiquitous DNA binding protein that promotes inflammatory tissue injury, is significantly elevated in CF sputum. However, its mechanistic and potential therapeutic implications in CF were previously unknown. We found that HMGB1 levels were significantly elevated in bronchoalveolar lavage fluids (BALs) of CF patients and cystic fibrosis transmembrane conductance regulator (CFTR)-/- mice. Neutralizing anti-HMGB1 monoclonal antibody (mAb) conferred significant protection against P aeruginosa-induced neutrophil recruitment, lung injury and bacterial infection in both CFTR-/- and wild-type mice. Alveolar macrophages isolated from mice treated with anti-HMGBl mAb had improved phagocytic activity, which was suppressed by direct exposure to HMGB1. In addition, BAL from CF patients significantly impaired macrophage phagocytotic function, and this impairment was attenuated by HMGB1-neutralizing antibodies. The HMGB1-mediated suppression of bacterial phagocytosis was attenuated in macrophages lacking toll-like receptor (TLR)-4, suggesting a critical role for TLR4 in signaling HMGB1-mediated macrophage dysfunction. These studies demonstrate that the elevated levels of HMGB1 in CF airways are critical for neutrophil recruitment and persistent presence of P. aeruginosa in the lung. Thus, HMGB1 may provide a therapeutic target for reducing bacterial infection and lung inflammation in CF.

Notes

Acknowledgments

This work was supported by grants (to LL Mantell) from the National Heart, Lung, and Blood Institute (NHLBI) (HL093708), St. John’s University and The Feinstein Institute for Medical Research, North Shore-Long Island Jewish Health System. The authors would like to thank Jenna L Bement, Koichiro Takahashi, Ashwini Gore and Vivek Patel for insightful discussions and excellent assistance. M Entezari’s current affiliation is LaGuardia Community College, City University of New York, New York, NY, USA.

References

  1. 1.
    Ratjen F, Doring G. (2003) Cystic fibrosis. Lancet. 361:681–9.CrossRefPubMedGoogle Scholar
  2. 2.
    Riordan JR, et al. (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 245:1066–73.CrossRefPubMedGoogle Scholar
  3. 3.
    Whittaker LA. (2007) Preface. Clin Chest Med. 28:xiii–xiv.CrossRefGoogle Scholar
  4. 4.
    Rowe SM, et al. (2008) Potential role of high-mobility group box 1 in cystic fibrosis airway disease. Am. J. Respir. Crit. Care Med. 178:822–31.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Høiby N, Koch C. (1990) Cystic fibrosis. 1. Pseudomonas aeruginosa infection in cystic fibrosis and its management. Thorax. 45:881–4.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Gibson RL, Burns JL, Ramsey BW. (2003) Pathophysiology and management of pulmonary infections in cystic fibrosis. Am. J. Respir. Crit. Care Med. 168:918–51.CrossRefPubMedGoogle Scholar
  7. 7.
    Brennan AL, Geddes DM. (2002) Cystic fibrosis. Curr. Opin. Infect. Dis. 15:175–82.CrossRefPubMedGoogle Scholar
  8. 8.
    Rosenfeld M, et al. (2001) Early pulmonary infection, inflammation, and clinical outcomes in infants with cystic fibrosis. Pediatr. Pulmonol. 32:356–66.CrossRefPubMedGoogle Scholar
  9. 9.
    Emerson J, Rosenfeld M, McNamara S, Ramsey B, Gibson RL. (2002) Pseudomonas aeruginosa and other predictors of mortality and morbidity in young children with cystic fibrosis. Pediatr. Pulmonol. 34:91–100.CrossRefPubMedGoogle Scholar
  10. 10.
    Pier GB, et al. (1996) Role of mutant CFTR in hypersusceptibility of cystic fibrosis patients to lung infections. Science. 271:64–7.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Pier GB, Grout M, Zaidi TS. (1997) Cystic fibrosis transmembrane conductance regulator is an epithelial cell receptor for clearance of Pseudomonas aeruginosa from the lung. Proc. Natl. Acad. Sci. U. S. A. 94:12088–93.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Greenberg S, Grinstein S. (2002) Phagocytosis and innate immunity. Curr. Opin. Immunol. 14:136–45.CrossRefPubMedGoogle Scholar
  13. 13.
    Brennan S, et al. (2009) Alveolar macrophages and CC chemokines are increased in children with cystic fibrosis. Eur. Respir. J. 34:655–61.CrossRefPubMedGoogle Scholar
  14. 14.
    Hubeau C, Puchelle E, Gaillard D. (2001) Distinct pattern of immune cell population in the lung of human fetuses with cystic fibrosis. J. Allergy Clin. Immunol. 108:524–9.CrossRefPubMedGoogle Scholar
  15. 15.
    Di A, et al. (2006) CFTR regulates phagosome acidification in macrophages and alters bactericidal activity. Nat. Cell Biol. 8:933–44.CrossRefPubMedGoogle Scholar
  16. 16.
    Thomassen MJ, et al. (1980) Ultrastructure and function of alveolar macrophages from cystic fibrosis patients. Pediatr. Res. 14:715–21.CrossRefPubMedGoogle Scholar
  17. 17.
    Wang H, et al. (1999) HMG-1 as a late mediator of endotoxin lethality in mice. Science. 285:248–51.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Abraham E, Arcaroli J, Carmody A, Wang H, Tracey KJ. (2000) Cutting edge: HMG-1 as a mediator of acute lung inflammation. J. Immunol. 165:2950–4.CrossRefPubMedGoogle Scholar
  19. 19.
    Sappington PL, et al. (2002) HMGB1 B box increases the permeability of Caco-2 enterocytic monolayers and impairs intestinal barrier function in mice. Gastroenterology. 123:790–802.CrossRefPubMedGoogle Scholar
  20. 20.
    Lin X, et al. (2005) Alpha-chemokine receptor blockade reduces high mobility group box 1 protein-induced lung inflammation and injury and improves survival in sepsis. Am. J. Physiol. Lung Cell. Mol. Physiol. 289:L583–90.CrossRefPubMedGoogle Scholar
  21. 21.
    Lotze MT, Tracey KJ. (2005) High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat. Rev. Immunol. 5:331–42.CrossRefPubMedGoogle Scholar
  22. 22.
    Liu G, et al. (2008) High mobility group protein-1 inhibits phagocytosis of apoptotic neutrophils through binding to phosphatidylserine. J. Immunol. 181:4240–6.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Gaggar A, Rowe SM, Hardison M, Blalock JE. (2010) Proline-glycine-proline (PGP) and high mobility group box protein-1 (HMGB1): potential mediators of cystic fibrosis airway inflammation. Open Respir. Med. J. 4:32–8.PubMedPubMedCentralGoogle Scholar
  24. 24.
    Qin S, et al. (2006) Role of HMGB1 in apoptosismediated sepsis lethality. J. Exp. Med. 203:1637–42.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Andersson U, et al. (2000) High mobility group 1 protein (HMG-1) stimulates proinflammatory cytokine synthesis in human monocytes. J. Exp. Med. 192:565–70.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Li J, et al. (2004) Recombinant HMGB1 with cytokine-stimulating activity. J. Immunol. Methods. 289:211–23.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Aida Y, Pabst MJ. (1990) Removal of endotoxin from protein solutions by phase separation using Triton X-114. J. Immunol. Methods. 132:191–5.CrossRefPubMedGoogle Scholar
  28. 28.
    Allard JB, et al. (2006) Aspergillus fumigatus generates an enhanced Th2-biased immune response in mice with defective cystic fibrosis transmembrane conductance regulator. J. Immunol. 177:5186–5194.CrossRefPubMedGoogle Scholar
  29. 29.
    Wargo MJ, Ho TC, Gross MJ, Whittaker LA, Hogan DA. (2009) GbdR regulates Pseudomonas aeruginosa plcH and pchP transcription in response to choline catabolites. Infect. Immun. 77:1103–11.CrossRefPubMedGoogle Scholar
  30. 30.
    Morrow DMP, et al. (2007) Antioxidants preserve macrophage phagocytosis of Pseudomonas aeruginosa during hyperoxia. 42:1338–49.Google Scholar
  31. 31.
    Mantell LL, et al. (1997) Unscheduled apoptosis during acute inflammatory lung injury. Cell Death Differ. 4:600–7.CrossRefPubMedGoogle Scholar
  32. 32.
    Yang H, et al. (2010) A critical cysteine is required for HMGB1 binding to Toll-like receptor 4 and activation of macrophage cytokine release. Proc. Natl. Acad. Sci. U. S. A. 107:11942–7.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Zhou L, et al. (1994) Correction of lethal intestinal defect in a mouse model of cystic fibrosis by human CFTR. Science. 266:1705–8.CrossRefPubMedGoogle Scholar
  34. 34.
    Loi R, Beckett T, Goncz KK, Suratt BT, Weiss DJ. (2006) Limited restoration of cystic fibrosis lung epithelium in vivo with adult bone marrow-derived cells. Am. J. Respir. Crit. Care Med. 173:171–9.CrossRefPubMedGoogle Scholar
  35. 35.
    Stover CK, et al. (2000) Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature. 406:959–64.CrossRefPubMedGoogle Scholar
  36. 36.
    Snouwaert JN, et al. (1992) An animal model for cystic fibrosis made by gene targeting. Science. 257:1083–8.CrossRefPubMedGoogle Scholar
  37. 37.
    Clarke LL, et al. (1992) Defective epithelial chloride transport in a gene-targeted mouse model of cystic fibrosis. Science. 257:1125–8.CrossRefPubMedGoogle Scholar
  38. 38.
    Burns JL, et al. (2001) Longitudinal assessment of Pseudomonas aeruginosa in young children with cystic fibrosis. J. Infect. Dis. 183:444–52.CrossRefPubMedGoogle Scholar
  39. 39.
    Yang H, et al. (2004) Reversing established sepsis with antagonists of endogenous high-mobility group box 1. Proc. Natl. Acad. Sci. U. S. A. 101:296–301.CrossRefPubMedGoogle Scholar
  40. 40.
    Khan TZ, et al. (1995) Early pulmonary inflammation in infants with cystic fibrosis. Am. J. Respir. Crit. Care Med. 151:1075–82.PubMedGoogle Scholar
  41. 41.
    Balough K, et al. (1995) The relationship between infection and inflammation in the early stages of lung disease from cystic fibrosis. Pediatr. Pulmonol. 20:63–70.CrossRefPubMedGoogle Scholar
  42. 42.
    Noah TL, Black HR, Cheng PW, Wood RE, Leigh MW. (1997) Nasal and bronchoalveolar lavage fluid cytokines in early cystic fibrosis. J. Infect. Dis. 175:638–47.CrossRefPubMedGoogle Scholar
  43. 43.
    Roum J, Buhl R, McElvaney N, Borok Z, Crystal R. (1993) Systemic deficiency of glutathione in cystic fibrosis. J. Appl. Physiol. 75:2419–24.CrossRefPubMedGoogle Scholar
  44. 44.
    Bonfield TL, Konstan MW, Berger M. (1999) Altered respiratory epithelial cell cytokine production in cystic fibrosis. J. Allergy Clin. Immunol. 104:72–8.CrossRefPubMedGoogle Scholar
  45. 45.
    Bonfield T, et al. (1995) Normal bronchial epithelial cells constitutively produce the anti-inflammatory cytokine interleukin-10, which is downregulated in cystic fibrosis. Am. J. Respir. Cell Mol. Biol. 13:257–61.CrossRefPubMedGoogle Scholar
  46. 46.
    Davis PB, Drumm M, Konstan MW. (1996) Cystic fibrosis. Am. J. Respir. Crit. Care Med. 154:1229–56.CrossRefPubMedGoogle Scholar
  47. 47.
    Knapp S, Schultz MJ, Poll T. (2005) Pneumonia models and innate immunity to respiratory bacterial pathogens. Shock. 24 Suppl 1:12–8.CrossRefPubMedGoogle Scholar
  48. 48.
    Zhang P, Summer WR, Bagby GJ, Nelson S. (2000) Innate immunity and pulmonary host defense. Immunol. Rev. 173:39–51.CrossRefPubMedGoogle Scholar
  49. 49.
    Harvey CJ, et al. (2011) Targeting Nrf2 signaling improves bacterial clearance by alveolar macrophages in patients with COPD and in a mouse model. Sci. Transl. Med. 3:78ra32.CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Reddy NM, et al. (2009) Innate immunity against bacterial infection following hyperoxia exposure is impaired in NRF2-deficient mice. J. Immunol. 183:4601–8.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Sun Y, et al. (2010) TLR4 and TLR5 on corneal macrophages regulate Pseudomonas aeruginosa keratitis by signaling through MyD88-dependent and -independent pathways. J. Immunol. 185:4272–83.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Thomassen MJ, et al. (1979) Inhibitory effect of cystic fibrosis serum on pseudomonas phagocytosis by rabbit and human alveolar macrophages. Pediatr. Res. 13:1085–8.CrossRefPubMedGoogle Scholar
  53. 53.
    Stoltz DA, et al. (2010) Cystic fibrosis pigs develop lung disease and exhibit defective bacterial eradication at birth. Sci. Transl. Med. 2:29ra31.CrossRefPubMedPubMedCentralGoogle Scholar
  54. 54.
    Calogero S, et al. (1999) The lack of chromosomal protein Hmg1 does not disrupt cell growth but causes lethal hypoglycaemia in newborn mice. Nat. Genet. 22:276–80.CrossRefPubMedGoogle Scholar
  55. 55.
    van Zoelen MAD, et al. (2009) Role of toll-like receptors 2 and 4, and the receptor for advanced glycation end products in high-mobility group box 1-induced inflammation in vivo. Shock. 31:280–4.CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Yu M, et al. (2006) HMGB1 signals through tolllike receptor (TLR) 4 and TLR2. Shock. 26:174–9.CrossRefPubMedGoogle Scholar
  57. 57.
    Park JS, et al. (2004) Involvement of toll-like receptors 2 and 4 in cellular activation by high mobility group box 1 protein. J. Biol. Chem. 279:7370–7.CrossRefPubMedGoogle Scholar
  58. 58.
    Park JS, et al. (2006) High mobility group box 1 protein interacts with multiple Toll-like receptors. Am. J. Physiol. Cell Physiol. 290:C917–24.CrossRefPubMedGoogle Scholar
  59. 59.
    Skerrett SJ, Wilson CB, Liggitt HD, Hajjar AM. (2007) Redundant Toll-like receptor signaling in the pulmonary host response to Pseudomonas aeruginosa. Am. J. Physiol. Lung Cell. Mol. Physiol. 292:L312–22.CrossRefPubMedGoogle Scholar
  60. 60.
    John G, Yildirim AO, Rubin BK, Gruenert DC, Henke MO. (2010) TLR-4-mediated innate immunity is reduced in cystic fibrosis airway cells. Am. J. Respir. Cell Mol. Biol. 42:424–31.CrossRefPubMedPubMedCentralGoogle 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 (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Maria Entezari
    • 1
  • Daniel J Weiss
    • 2
  • Ravikumar Sitapara
    • 1
  • Laurie Whittaker
    • 2
  • Matthew J Wargo
    • 2
  • JianHua Li
    • 3
  • Haichao Wang
    • 3
  • Huan Yang
    • 3
  • Lokesh Sharma
    • 1
  • Binh D Phan
    • 1
  • Mohammad Javdan
    • 3
  • Sangeeta S Chavan
    • 3
  • Edmund J Miller
    • 4
  • Kevin J Tracey
    • 3
  • Lin L Mantell
    • 1
    • 3
    • 4
  1. 1.Cardiopulmonary Toxicology, Department of Pharmaceutical SciencesSt. John’s University College of Pharmacy and Allied Health ProfessionsQueensUSA
  2. 2.Pulmonary and Critical CareUniversity of Vermont College of MedicineBurlingtonUSA
  3. 3.Inflammation and ImmunologyThe Feinstein Institute for Medical ResearchManhassetUSA
  4. 4.Heart and Lung ResearchThe Feinstein Institute for Medical ResearchManhassetUSA

Personalised recommendations