Molecular Medicine

, Volume 16, Issue 9–10, pp 343–351 | Cite as

Immunomodulatory Drugs Regulate HMGB1 Release from Activated Human Monocytes

  • Hanna Schierbeck
  • Heidi Wähämaa
  • Ulf Andersson
  • Helena Erlandsson Harris
Research Article


Several HMGBl-specific antagonists have provided beneficial results in multiple models of inflammatory disease-preclinical trials including arthritis. Since no HMGB1-specific targeted therapy has yet reached the clinic, we have performed in vitro studies to investigate whether any of a selection of well-established antirheumatic drugs inhibit HMGB1 release as part of its mode of action. Freshly purified peripheral blood monocytes from healthy donors were stimulated in cultures with LPS and IFNγ to cause HMGB1 and TNF release detected in ELISPOT assays. Effects on the secretion were assessed in cultures supplemented with dexamethasone, cortisone, chloroquine, gold sodium thiomalate, methotrexate, colchicine, etanercept or anakinra. Pharmacologically relevant doses of dexamethasone, gold sodium thiomalate and chloroquine inhibited the extracellular release of HMGB1 in a dose-dependent mode. Immunostaining demonstrated that dexamethasone caused intracellular HMGB1 retention. No effects on HMGB1 secretion were observed in cultures with activated monocytes by any of the other studied agents. TNF production in LPS/IFNγ-activated monocytes was readily downregulated by dexamethasone and, to some extent, by chloroquine and etanercept. We conclude that dexamethasone, gold sodium thiomalate and chloroquine share a capacity to inhibit HMGB1 release from activated monocytes.



Financial support was provided through the regional agreement on medical training and clinical research (ALF) between Stockholm County Council and Karolinska Institutet, the Swedish Association against Rheumatism, the Swedish Medical Research Council, the Freemason Lodge Barnhuset in Stockholm, and King Gustaf V’s Foundation.


  1. 1.
    Yang H, Tracey KJ. Targeting HMGB1 in inflammation. Biochim. Biophys. Acta. 1799:149–56.Google Scholar
  2. 2.
    Andersson U, Harris HE. The role of HMGB1 in the pathogenesis of rheumatic disease. Biochim. Biophys. Acta. 1799:141–8.Google Scholar
  3. 3.
    Semino C, Angelini G, Poggi A, Rubartelli A. (2005) NK/iDC interaction results in IL-18 secretion by DCs at the synaptic cleft followed by NK cell activation and release of the DC maturation factor HMGB1. Blood. 106:609–16.CrossRefGoogle Scholar
  4. 4.
    Wang H, et al. (1999) HMG-1 as a late mediator of endotoxin lethality in mice. Science. 285:248–51.CrossRefGoogle Scholar
  5. 5.
    Wang H, et al. (1999) Proinflammatory cytokines (tumor necrosis factor and interleukin 1) stimulate release of high mobility group protein-1 by pituicytes. Surgery. 126:389–92.CrossRefGoogle Scholar
  6. 6.
    Rendon-Mitchell B, et al. (2003) IFN-gamma induces high mobility group box 1 protein release partly through a TNF-dependent mechanism. J. Immunol. 170:3890–7.CrossRefGoogle Scholar
  7. 7.
    Tang D, et al. (2007) The anti-inflammatory effects of heat shock protein 72 involve inhibition of high-mobility-group box 1 release and proinflammatory function in macrophages. J. Immunol. 179:1236–44.CrossRefGoogle Scholar
  8. 8.
    Oh YJ, et al. (2009) HMGB1 is phosphorylated by classical protein kinase C and is secreted by a calcium-dependent mechanism. J. Immunol. 182:5800–9.CrossRefGoogle Scholar
  9. 9.
    Kawahara K, et al. (2008) C-reactive protein induces high-mobility group box-1 protein release through activation of p38MAPK in macrophage RAW264.7 cells. Cardiovasc. Pathol. 17:129–38.CrossRefGoogle Scholar
  10. 10.
    Youn JH, Shin JS. (2006) Nucleocytoplasmic shuttling of HMGB1 is regulated by phosphorylation that redirects it toward secretion. J. Immunol. 177:7889–97.CrossRefGoogle Scholar
  11. 11.
    Bonaldi T, et al. (2003) Monocytic cells hyper-acetylate chromatin protein HMGB1 to redirect it towards secretion. Embo. J. 22:5551–60.CrossRefGoogle Scholar
  12. 12.
    Gardella S, et al. (2002) The nuclear protein HMGB1 is secreted by monocytes via a non-classical, vesicle-mediated secretory pathway. EMBO Rep. 3:995–1001.CrossRefGoogle Scholar
  13. 13.
    Harris HE, Raucci A. (2006) Alarmin(g) news about danger: workshop on innate danger signals and HMGB1. EMBO Rep. 7:774–8.PubMedPubMedCentralGoogle Scholar
  14. 14.
    Touqui L, Alaoui-El-Azher M. (2001) Mammalian secreted phospholipases A2 and their pathophysiological significance in inflammatory diseases. Curr. Mol. Med. 1:739–54.CrossRefGoogle Scholar
  15. 15.
    Scaffidi P, Misteli T, Bianchi ME. (2002) Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature. 418:191–5.CrossRefGoogle Scholar
  16. 16.
    Bell CW, Jiang W, Reich CF 3rd, Pisetsky DS. (2006) The extracellular release of HMGB1 during apoptotic cell death. Am. J. Physiol. Cell Physiol. 291:C1318–25.CrossRefGoogle Scholar
  17. 17.
    Schroder K, Hertzog PJ, Ravasi T, Hume DA. (2004) Interferon-gamma: an overview of signals, mechanisms and functions. J. Leukoc. Biol. 75:163–89.CrossRefGoogle Scholar
  18. 18.
    Wahamaa H, et al. (2007) HMGB1-secreting capacity of multiple cell lineages revealed by a novel HMGB1 ELISPOT assay. J. Leukoc. Biol. 81:129–36.CrossRefGoogle Scholar
  19. 19.
    Lotze MT, Tracey KJ. (2005) High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat. Rev. Immunol. 5:331–42.CrossRefGoogle Scholar
  20. 20.
    Czock D, Keller F, Rasche FM, Haussler U. (2005) Pharmacokinetics and pharmacodynamics of systemically administered glucocorticoids. Clin. Pharmacokinet. 44:61–98.CrossRefGoogle Scholar
  21. 21.
    Shiozawa K, et al. (2005) Serum levels and pharmacodynamics of methotrexate and its metabolite 7-hydroxy methotrexate in Japanese patients with rheumatoid arthritis treated with 2-mg capsule of methotrexate three times per week. Mod. Rheumatol. 15:405–9.CrossRefGoogle Scholar
  22. 22.
    Korth-Bradley JM, Rubin AS, Hanna RK, Simcoe DK, Lebsack ME. (2000) The pharmacokinetics of etanercept in healthy volunteers. Ann. Pharmacother. 34:161–4.CrossRefGoogle Scholar
  23. 23.
    Granowitz EV, et al. (1992) Pharmacokinetics, safety and immunomodulatory effects of human recombinant interleukin-1 receptor antagonist in healthy humans. Cytokine. 4:353–60.CrossRefGoogle Scholar
  24. 24.
    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.CrossRefGoogle Scholar
  25. 25.
    Abraham E, Arcaroli J, Carmody A, Wang H, Tracey KJ. (2000) HMG-1 as a mediator of acute lung inflammation. Immunol. 165:2950–1.CrossRefGoogle Scholar
  26. 26.
    Kim JY, et al. (2005) HMGB1 contributes to the development of acute lung injury after hemorrhage. Am. J. Physiol. Lung Cell. Mol. Physiol. 288:L958–65.CrossRefGoogle Scholar
  27. 27.
    Liu K, et al. (2007) Anti-high mobility group box 1 monoclonal antibody ameliorates brain infarction induced by transient ischemia in rats. Faseb. J. 21:3904–16.CrossRefGoogle Scholar
  28. 28.
    Van de Wouwer M, et al. (2006) The lectin-like domain of thrombomodulin interferes with complement activation and protects against arthritis. J. Thromb. Haemost. 4:1813–24.CrossRefGoogle Scholar
  29. 29.
    Hofmann MA, et al. (2002) RAGE and arthritis: the G82S polymorphism amplifies the inflammatory response. Genes Immun. 3:123–35.CrossRefGoogle Scholar
  30. 30.
    Kokkola R, et al. (2003) Successful treatment of collagen-induced arthritis in mice and rats by targeting extracellular high mobility group box chromosomal protein 1 activity. Arthritis Rheum. 48:2052–8.CrossRefGoogle Scholar
  31. 31.
    Lutterloh EC, et al. (2007) Inhibition of the RAGE products increases survival in experimental models of severe sepsis and systemic infection. Crit. Care. 11:R122.CrossRefGoogle Scholar
  32. 32.
    Huston JM, et al. (2007) Transcutaneous vagus nerve stimulation reduces serum high mobility group box 1 levels and improves survival in murine sepsis. Crit. Care Med. 35:2762–8.PubMedGoogle Scholar
  33. 33.
    Pan P, et al. (2009) Low-dose cisplatin administration in murine cecal ligation and puncture prevents the systemic release of HMGB1 and attenuates lethality. J. Leukoc. Biol. 86:625–32.CrossRefGoogle Scholar
  34. 34.
    Ostberg T, et al. (2008) Oxaliplatin retains HMGB1 intranuclearly and ameliorates collagen type II-induced arthritis. Arthritis Res. Ther. 10: R1.CrossRefGoogle Scholar
  35. 35.
    Klint E, et al. (2005) Intraarticular glucocorticoid treatment reduces inflammation in synovial cell infiltrations more efficiently than in synovial blood vessels. Arthritis Rheum. 52:3880–9.CrossRefGoogle Scholar
  36. 36.
    Thieringer R, et al. (2001) 11 Beta-hydroxysteroid dehydrogenase type 1 is induced in human monocytes upon differentiation to macrophages. J. Immunol. 167:30–5.CrossRefGoogle Scholar
  37. 37.
    Zetterstrom CK, et al. (2008) Pivotal advance: inhibition of HMGB1 nuclear translocation as a mechanism for the anti-rheumatic effects of gold sodium thiomalate. J. Leukoc. Biol. 83:31–8.CrossRefGoogle Scholar
  38. 38.
    French JK, Hurst NP, O’Donnell ML, Betts WH. (1987) Uptake of chloroquine and hydroxy-chloroquine by human blood leucocytes in vitro: relation to cellular concentrations during antirheumatic therapy. Ann. Rheum. Dis. 46:42–5.CrossRefGoogle Scholar
  39. 39.
    Tett S, Cutler D, Day R. (1990) Antimalarials in rheumatic diseases. Baillieres Clin. Rheumatol. 4:467–89.CrossRefGoogle Scholar
  40. 40.
    Lange F, et al. (2005) Methotrexate ameliorates T cell dependent autoimmune arthritis and encephalomyelitis but not antibody induced or fibroblast induced arthritis. Ann. Rheum. Dis. 64:599–605.CrossRefGoogle Scholar
  41. 41.
    Sundberg E, et al. (2008) Systemic TNF blockade does not modulate synovial expression of the pro-inflammatory mediator HMGB1 in rheumatoid arthritis patientsa prospective clinical study. Arthritis Res. Ther. 10:R33.CrossRefGoogle Scholar
  42. 42.
    Sha Y, Zmijewski J, Xu Z, Abraham E. (2008) HMGB1 develops enhanced proinflammatory activity by binding to cytokines. J. Immunol. 180:2531–7.CrossRefGoogle Scholar

Copyright information

© The Feinstein Institute for Medical Research 2010

Authors and Affiliations

  • Hanna Schierbeck
    • 1
    • 3
  • Heidi Wähämaa
    • 1
    • 3
  • Ulf Andersson
    • 1
    • 3
  • Helena Erlandsson Harris
    • 2
    • 3
  1. 1.Department of Women’s and Children’s Health, Pediatric UnitKarolinska Institutet/Karolinska University Hospital Q1:02StockholmSweden
  2. 2.Department of Medicine, Rheumatology UnitKarolinska InstitutetStockholmSweden
  3. 3.Rheumatology Research Laboratory, Center for Molecular Medicine, Karolinska InstitutetKarolinska University HospitalStockholmSweden

Personalised recommendations