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Extracellular histones are major mediators of death in sepsis

Nature Medicine volume 15, pages 1318–1321 (2009)Cite this article

Abstract

Hyperinflammatory responses can lead to a variety of diseases, including sepsis1. We now report that extracellular histones released in response to inflammatory challenge contribute to endothelial dysfunction, organ failure and death during sepsis. They can be targeted pharmacologically by antibody to histone or by activated protein C (APC). Antibody to histone reduced the mortality of mice in lipopolysaccharide (LPS), tumor necrosis factor (TNF) or cecal ligation and puncture models of sepsis. Extracellular histones are cytotoxic toward endothelium in vitro and are lethal in mice. In vivo, histone administration resulted in neutrophil margination, vacuolated endothelium, intra-alveolar hemorrhage and macro- and microvascular thrombosis. We detected histone in the circulation of baboons challenged with Escherichia coli, and the increase in histone levels was accompanied by the onset of renal dysfunction. APC cleaves histones and reduces their cytotoxicity. Co-infusion of APC with E. coli in baboons or histones in mice prevented lethality. Blockade of protein C activation exacerbated sublethal LPS challenge into lethality, which was reversed by treatment with antibody to histone. We conclude that extracellular histones are potential molecular targets for therapeutics for sepsis and other inflammatory diseases.

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Figure 1: Cytotoxicity of extracellular histones toward endothelium and APC cleavage of histones.
Figure 2: APC cleaves histones both in vitro and in vivo.
Figure 3: Intravenous injection of histones elicits inflammatory and cell injury responses.
Figure 4: Antibody to H4 protects mice from the lethality of LPS, CLP and TNF in vivo.

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Acknowledgements

We thank K. Jackson for protein sequencing and identification; A. Matveev for preparing liposomes; C.J. Edgell, University of North Carolina, Chapel Hill, for providing EA.hy926 cells; G. Kinasewitz, University of Oklahoma Health Science Center, for providing plasma samples from humans with sepsis; K. Deatherage for preparing the manuscript; and P. Kincade for reviewing the original manuscript. C.T.E. is an investigator of the Howard Hughes Medical Institute. F.L. and F.B.T. are supported by US National Institutes of Health grant R01GM037704. C.T.A. and F.S. are supported by fellowships from the University of Bari, Italy.

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Authors and Affiliations

  1. Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA

    Jun Xu, Concetta T Ammollo, Fabrizio Semeraro, Fletcher B Taylor, Naomi L Esmon, Florea Lupu & Charles T Esmon

  2. Howard Hughes Medical Institute, Oklahoma City, Oklahoma, USA

    Xiaomei Zhang & Charles T Esmon

  3. Immunology and Cancer Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA

    Rosana Pelayo

  4. Department of Microbiology and Immunology, Temple Autoimmunity Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA

    Marc Monestier

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  1. Jun Xu
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  2. Xiaomei Zhang
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  8. Naomi L Esmon
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Contributions

J.X. designed and executed many of the experiments; X.Z. participated in the experimental execution and contributed to experimental design; R.P. was involved in the initial experiments to activate the macrophages, leading to the release of histones; M.M. provided the antibodies to the histones and contributed useful comments; C.T.A. and F.S. performed the CLP experiments; F.B.T. provided the archival baboon sepsis plasma and contributed useful comments; N.L.E. made useful comments and assisted in manuscript preparation; F.L. provided constructive critiques of the studies and performed the calcium flux experiments and the histochemical analysis of the tissues; and C.T.E. oversaw the overall execution of the projects.

Corresponding author

Correspondence to Charles T Esmon.

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Supplementary Figs. 1–4 and Supplementary Methods (PDF 402 kb)

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Xu, J., Zhang, X., Pelayo, R. et al. Extracellular histones are major mediators of death in sepsis. Nat Med 15, 1318–1321 (2009). https://doi.org/10.1038/nm.2053

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