Skip to main content
SpringerLink
Log in

Analysis of the Released Nuclear Cytokine HMGB1 in Human Serum

  • Protocol
  • First Online:
Cytokine Bioassays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1172))

Abstract

A ubiquitous nuclear protein, the high-mobility group box 1 (HMGB1), is secreted by activated macrophages/monocytes and leaked passively from injured cells. HMGB1 functions as a mediator of infection- and injury-elicited inflammatory diseases. Here, we describe a semiquantitative immuno-blotting method to measure the released HMGB1 in human serum, in comparison with a commercially available HMGB1 ELISA technique.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Wang H, Bloom O, Zhang M et al (1999) HMG-1 as a late mediator of endotoxin lethality in mice. Science 285:248–251

    Article  CAS  PubMed  Google Scholar 

  2. Bonaldi T, Talamo F, Scaffidi P et al (2003) Monocytic cells hyperacetylate chromatin protein HMGB1 to redirect it towards secretion. EMBO J 22:5551–5560

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Lotze MT, Tracey KJ (2005) High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol 5:331–342

    Article  CAS  PubMed  Google Scholar 

  4. Ivanov S, Dragoi AM, Wang X et al (2007) A novel role for HMGB1 in TLR9-mediated inflammatory responses to CpG-DNA. Blood 110:1970–1981

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Rendon-Mitchell B, Ochani M, Li J et al (2003) IFN-gamma induces high mobility group box 1 protein release partly through a TNF-dependent mechanism. J Immunol 170:3890–3897

    Article  CAS  PubMed  Google Scholar 

  6. Tang D, Shi Y, Kang R et al (2007) Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB1. J Leukoc Biol 81:741–747

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Scaffidi P, Misteli T, Bianchi ME (2002) Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 418:191–195

    Article  CAS  PubMed  Google Scholar 

  8. Wang H, Ward MF, Fan XG et al (2006) Potential role of high mobility group box 1 in viral infectious diseases. Viral Immunol 19:3–9

    Article  PubMed Central  PubMed  Google Scholar 

  9. Zhu S, Li W, Ward MF et al (2010) High mobility group box 1 protein as a potential drug target for infection- and injury-elicited inflammation. Inflamm Allergy Drug Targets 9:60–72

    Article  PubMed Central  PubMed  Google Scholar 

  10. Wang H, Ward MF, Sama AE (2009) Novel HMGB1-inhibiting therapeutic agents for experimental sepsis. Shock 32:348–357

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Wang H, Yang H, Tracey KJ (2004) Extracellular role of HMGB1 in inflammation and sepsis. J Intern Med 255:320–331

    Article  CAS  PubMed  Google Scholar 

  12. Wang H, Li W, Goldstein R et al (2007) HMGB1 as a potential therapeutic target. Novartis Found Symp 280:73–85

    Article  CAS  PubMed  Google Scholar 

  13. Wang H, Zhu S, Zhou R et al (2008) Therapeutic potential of HMGB1-targeting agents in sepsis. Expert Rev Mol Med 10:e32

    Article  PubMed Central  PubMed  Google Scholar 

  14. Urbonaviciute V, Furnrohr BG, Weber C et al (2007) Factors masking HMGB1 in human serum and plasma. J Leukoc Biol 81:67–74

    Article  CAS  PubMed  Google Scholar 

  15. Sha Y, Zmijewski J, Xu Z et al (2008) HMGB1 develops enhanced proinflammatory activity by binding to cytokines. J Immunol 180:2531–2537

    Article  CAS  PubMed  Google Scholar 

  16. He RL, Zhou J, Hanson CZ et al (2009) Serum amyloid A induces G-CSF expression and neutrophilia via Toll-like receptor 2. Blood 113:429–437

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Lehner J, Wittwer C, Fersching D et al (2012) Methodological and preanalytical evaluation of an HMGB1 immunoassay. Anticancer Res 32:2059–2062

    CAS  PubMed  Google Scholar 

  18. Yasuda T, Ueda T, Takeyama Y et al (2006) Significant increase of serum high-mobility group box chromosomal protein 1 levels in patients with severe acute pancreatitis. Pancreas 33:359–363

    Article  CAS  PubMed  Google Scholar 

  19. Gaini S, Pedersen SS, Koldkjaer OG et al (2007) High mobility group box-1 protein in patients with suspected community-acquired infections and sepsis: a prospective study. Crit Care 11:R32

    Article  PubMed Central  PubMed  Google Scholar 

  20. Okazaki K, Kondo M, Kato M et al (2008) Elevation of high-mobility group box 1 concentration in asphyxiated neonates. Neonatology 94:105–109

    Article  CAS  PubMed  Google Scholar 

  21. Karlsson S, Pettila V, Tenhunen J et al (2008) HMGB1 as a predictor of organ dysfunction and outcome in patients with severe sepsis. Intensive Care Med 34:1046–1053

    Article  CAS  PubMed  Google Scholar 

  22. Rowe SM, Jackson PL, Liu G et al (2008) Potential role of high-mobility group box 1 in cystic fibrosis airway disease. Am J Respir Crit Care Med 178:822–831

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. Yoshizaki A, Komura K, Iwata Y et al (2009) Clinical significance of serum HMGB-1 and sRAGE levels in systemic sclerosis: association with disease severity. J Clin Immunol 29:180–189

    Article  CAS  PubMed  Google Scholar 

  24. Chung HW, Lee SG, Kim H et al (2009) Serum high mobility group box-1 (HMGB1) is closely associated with the clinical and pathologic features of gastric cancer. J Transl Med 7:38. doi:10.1186/1479-5876-7-38

    Article  PubMed Central  PubMed  Google Scholar 

  25. Dupire G, Nicaise C, Gangji V et al (2012) Increased serum levels of high-mobility group box 1 (HMGB1) in primary Sjogren's syndrome. Scand J Rheumatol 41:120–123

    Article  CAS  PubMed  Google Scholar 

  26. Lee H, Song M, Shin N et al (2012) Diagnostic significance of serum HMGB1 in colorectal carcinomas. PLoS One 7:e34318

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Wang H, Yang H, Czura CJ et al (2001) HMGB1 as a late mediator of lethal systemic inflammation. Am J Respir Crit Care Med 164:1768–1773

    Article  CAS  PubMed  Google Scholar 

  28. Angus DC, Yang L, Kong L et al (2007) Circulating high-mobility group box 1 (HMGB1) concentrations are elevated in both uncomplicated pneumonia and pneumonia with severe sepsis. Crit Care Med 35:1061–1067

    Article  PubMed  Google Scholar 

  29. Kazama H, Ricci JE, Herndon JM et al (2008) Induction of immunological tolerance by apoptotic cells requires caspase-dependent oxidation of high-mobility group box-1 protein. Immunity 29:21–32

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  30. Park JS, Arcaroli J, Yum HK et al (2003) Activation of gene expression in human neutrophils by high mobility group box 1 protein. Am J Physiol Cell Physiol 284:C870–C879

    Article  CAS  PubMed  Google Scholar 

  31. Li W, Li J, Sama AE et al (2013) Carbenoxolone blocks endotoxin-induced PKR activation and HMGB1 release. Mol Med 19:203–211

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  32. Li W, Li J, Ashok M et al (2007) A cardiovascular drug rescues mice from lethal sepsis by selectively attenuating a late-acting proinflammatory mediator, high mobility group box 1. J Immunol 178:3856–3864

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  33. Wang H, Liao H, Ochani M et al (2004) Cholinergic agonists inhibit HMGB1 release and improve survival in experimental sepsis. Nat Med 10:1216–1221

    Article  CAS  PubMed  Google Scholar 

  34. Li W, Zhu S, Li J et al (2011) A hepatic protein, fetuin-A, occupies a protective role in lethal systemic inflammation. PLoS One 6:e16945

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Li W, Ashok M, Li J et al (2007) A major ingredient of green tea rescues mice from lethal sepsis partly by inhibiting HMGB1. PLoS One 2:e1153

    Article  PubMed Central  PubMed  Google Scholar 

  36. Yang H, Ochani M, Li J 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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  37. Manganelli V, Signore M, Pacini I et al (2010) Increased HMGB1 expression and release by mononuclear cells following surgical/anesthesia trauma. Crit Care 14:R197

    Article  PubMed Central  PubMed  Google Scholar 

  38. Zhou Y, Xiong KL, Lin S et al (2010) Elevation of high-mobility group protein box-1 in serum correlates with severity of acute intracerebral hemorrhage. Mediators Inflamm pii: 142458. doi: 10.1155/2010/142458

  39. Bruchfeld A, Wendt M, Bratt J et al (2011) High-mobility group box-1 protein (HMGB1) is increased in antineutrophilic cytoplasmatic antibody (ANCA)-associated vasculitis with renal manifestations. Mol Med 17:29–35

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  40. Shang GH, Jia CQ, Tian H et al (2009) Serum high mobility group box protein 1 as a clinical marker for non-small cell lung cancer. Respir Med 103:1949–1953

    Article  PubMed  Google Scholar 

  41. Goldstein RS, Gallowitsch-Puerta M, Yang L et al (2006) Elevated high-mobility group box 1 levels in patients with cerebral and myocardial ischemia. Shock 25:571–574

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments and Funding

Work in the authors’ laboratory was supported by grants from the National Institutes of Health (R01GM063075 and R01AT05076).

Author information

Authors and Affiliations

  1. Laboratory of Emergency Medicine, The Feinstein Institute for Medical Research, North Shore-LIJ Health System, 350 Community Drive, 11030, Manhasset, NY, USA

    Haichao Wang, Lin Zhao, Shu Zhu & Maggie Yeung

  2. Laboratory of Biomedical Sciences, The Feinstein Institute for Medical Research, North Shore-LIJ Health System, 350 Community Drive, 11030, Manhasset, NY, USA

    Jianhua Li

Authors
  1. Haichao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lin Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianhua Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shu Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maggie Yeung
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Haichao Wang .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, St. John's University, New York, New York, USA

    Ivana Vancurova

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, New York

About this protocol

Cite this protocol

Wang, H., Zhao, L., Li, J., Zhu, S., Yeung, M. (2014). Analysis of the Released Nuclear Cytokine HMGB1 in Human Serum. In: Vancurova, I. (eds) Cytokine Bioassays. Methods in Molecular Biology, vol 1172. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0928-5_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0928-5_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0927-8

  • Online ISBN: 978-1-4939-0928-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation