{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T16:43:38Z","timestamp":1793119418987,"version":"4.5.0"},"reference-count":29,"publisher":"Rockefeller University Press","issue":"1","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2015,1,12]]},"abstract":"<jats:p>Innate immune receptors for pathogen- and damage-associated molecular patterns (PAMPs and DAMPs) orchestrate inflammatory responses to infection and injury. Secreted by activated immune cells or passively released by damaged cells, HMGB1 is subjected to redox modification that distinctly influences its extracellular functions. Previously, it was unknown how the TLR4 signalosome distinguished between HMGB1 isoforms. Here we demonstrate that the extracellular TLR4 adaptor, myeloid differentiation factor 2 (MD-2), binds specifically to the cytokine-inducing disulfide isoform of HMGB1, to the exclusion of other isoforms. Using MD-2\u2013deficient mice, as well as MD-2 silencing in macrophages, we show a requirement for HMGB1-dependent TLR4 signaling. By screening HMGB1 peptide libraries, we identified a tetramer (FSSE, designated P5779) as a specific MD-2 antagonist preventing MD-2\u2013HMGB1 interaction and TLR4 signaling. P5779 does not interfere with lipopolysaccharide-induced cytokine\/chemokine production, thus preserving PAMP-mediated TLR4\u2013MD-2 responses. Furthermore, P5779 can protect mice against hepatic ischemia\/reperfusion injury, chemical toxicity, and sepsis. These findings reveal a novel mechanism by which innate systems selectively recognize specific HMGB1 isoforms. The results may direct toward strategies aimed at attenuating DAMP-mediated inflammation while preserving antimicrobial immune responsiveness.<\/jats:p>","DOI":"10.1084\/jem.20141318","type":"journal-article","created":{"date-parts":[[2015,1,5]],"date-time":"2015-01-05T22:41:16Z","timestamp":1420497676000},"page":"5-14","update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":362,"title":["MD-2 is required for disulfide HMGB1\u2013dependent TLR4 signaling"],"prefix":"10.1084","volume":"212","author":[{"given":"Huan","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haichao","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, North Shore University Hospital, Manhasset, NY 11030 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongliang","family":"Ju","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed A.","family":"Ragab","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Lundb\u00e4ck","sequence":"additional","affiliation":[{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"},{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Long","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, North Shore University Hospital, Manhasset, NY 11030 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergio I.","family":"Valdes-Ferrer","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingzhu","family":"He","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John P.","family":"Pribis","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213 8"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhua","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ben","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Domokos","family":"Gero","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, The University of Texas Medical Branch, Galveston, TX 77555 9"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csaba","family":"Szabo","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, The University of Texas Medical Branch, Galveston, TX 77555 9"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel J.","family":"Antoine","sequence":"additional","affiliation":[{"name":"Medical Research Council Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, University of Liverpool, Liverpool L69 3BX, England, UK 6"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helena E.","family":"Harris","sequence":"additional","affiliation":[{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"},{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doug T.","family":"Golenbock","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, MA 01655 7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianmin","family":"Meng","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, MA 01655 7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesse","family":"Roth","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sangeeta S.","family":"Chavan","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulf","family":"Andersson","sequence":"additional","affiliation":[{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"},{"name":"Department of Medicine 4 and 5"},{"name":"Department of Women\u2019s and Children\u2019s Health, Karolinska Institutet, Karolinska University Hospital, SE-171 77 Stockholm, Sweden 4 and 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy R.","family":"Billiar","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213 8"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin J.","family":"Tracey","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yousef","family":"Al-Abed","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science 1 and 2"},{"name":"Department of Medicinal Chemistry, The Feinstein Institute for Medical Research, Manhasset, NY 11030 1 and 2"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2015,1,5]]},"reference":[{"key":"2023072602431749600_bib1","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1146\/annurev-immunol-030409-101323","article-title":"HMGB1 is a therapeutic target for sterile inflammation and infection","volume":"29","author":"Andersson","year":"2011","journal-title":"Annu. Rev. Immunol."},{"key":"2023072602431749600_bib2","doi-asserted-by":"publisher","first-page":"1070","DOI":"10.1016\/j.jhep.2011.12.019","article-title":"Molecular forms of HMGB1 and keratin-18 as mechanistic biomarkers for mode of cell death and prognosis during clinical acetaminophen hepatotoxicity","volume":"56","author":"Antoine","year":"2012","journal-title":"J. Hepatol."},{"key":"2023072602431749600_bib3","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1002\/hep.26294","article-title":"Mechanistic biomarkers provide early and sensitive detection of acetaminophen-induced acute liver injury at first presentation to hospital","volume":"58","author":"Antoine","year":"2013","journal-title":"Hepatology."},{"key":"2023072602431749600_bib4","doi-asserted-by":"publisher","first-page":"1722","DOI":"10.1126\/science.1168988","article-title":"CD24 and Siglec-10 selectively repress tissue damage-induced immune responses","volume":"323","author":"Chen","year":"2009","journal-title":"Science."},{"key":"2023072602431749600_bib5","doi-asserted-by":"publisher","first-page":"5152","DOI":"10.4049\/jimmunol.1300496","article-title":"Lipopolysaccharide clearance, bacterial clearance, and systemic inflammatory responses are regulated by cell type-specific functions of TLR4 during sepsis","volume":"190","author":"Deng","year":"2013","journal-title":"J. Immunol."},{"key":"2023072602431749600_bib6","doi-asserted-by":"publisher","first-page":"382","DOI":"10.1016\/S0168-8278(03)00005-9","article-title":"Knodell RG, Ishak KG, Black WC, Chen TS, Craig R, Kaplowitz N, Kiernan TW, Wollman J. Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis [Hepatology 1981;1:431-435]","volume":"38","author":"Desmet","year":"2003","journal-title":"J. Hepatol."},{"key":"2023072602431749600_bib7","doi-asserted-by":"publisher","first-page":"19919","DOI":"10.1074\/jbc.274.28.19919","article-title":"Receptor for advanced glycation end products (RAGE)-mediated neurite outgrowth and activation of NF-\u03baB require the cytoplasmic domain of the receptor but different downstream signaling pathways","volume":"274","author":"Huttunen","year":"1999","journal-title":"J. Biol. Chem."},{"key":"2023072602431749600_bib8","doi-asserted-by":"publisher","first-page":"8946","DOI":"10.1128\/JVI.01857-08","article-title":"Toll-like receptor 4 deficiency increases disease and mortality after mouse hepatitis virus type 1 infection of susceptible C3H mice","volume":"83","author":"Khanolkar","year":"2009","journal-title":"J. Virol."},{"key":"2023072602431749600_bib9","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1016\/j.jim.2004.04.019","article-title":"Recombinant HMGB1 with cytokine-stimulating activity","volume":"289","author":"Li","year":"2004","journal-title":"J. Immunol. Methods."},{"key":"2023072602431749600_bib10","doi-asserted-by":"publisher","first-page":"8695","DOI":"10.1074\/jbc.M109.075127","article-title":"MD-2-mediated ionic interactions between lipid A and TLR4 are essential for receptor activation","volume":"285","author":"Meng","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2023072602431749600_bib11","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1038\/ni809","article-title":"Essential role of MD-2 in LPS responsiveness and TLR4 distribution","volume":"3","author":"Nagai","year":"2002","journal-title":"Nat. Immunol."},{"key":"2023072602431749600_bib12","doi-asserted-by":"publisher","first-page":"1191","DOI":"10.1038\/nature07830","article-title":"The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex","volume":"458","author":"Park","year":"2009","journal-title":"Nature."},{"key":"2023072602431749600_bib13","doi-asserted-by":"publisher","first-page":"1637","DOI":"10.1084\/jem.20052203","article-title":"Role of HMGB1 in apoptosis-mediated sepsis lethality","volume":"203","author":"Qin","year":"2006","journal-title":"J. Exp. Med."},{"key":"2023072602431749600_bib14","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1038\/nature00858","article-title":"Release of chromatin protein HMGB1 by necrotic cells triggers inflammation","volume":"418","author":"Scaffidi","year":"2002","journal-title":"Nature."},{"key":"2023072602431749600_bib15","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1097\/SHK.0b013e318276f4ca","article-title":"Plasma bacterial and mitochondrial DNA distinguish bacterial sepsis from sterile systemic inflammatory response syndrome and quantify inflammatory tissue injury in nonhuman primates","volume":"39","author":"Sursal","year":"2013","journal-title":"Shock."},{"key":"2023072602431749600_bib16","doi-asserted-by":"publisher","first-page":"1360","DOI":"10.2119\/molmed.2012.00314","article-title":"A Janus tale of two active high mobility group box 1 (HMGB1) redox states","volume":"18","author":"Tang","year":"2012","journal-title":"Mol. Med."},{"key":"2023072602431749600_bib17","doi-asserted-by":"publisher","first-page":"21881","DOI":"10.1074\/jbc.M800672200","article-title":"Isolation of monomeric and dimeric secreted MD-2. Endotoxin\u00b7sCD14 and Toll-like receptor 4 ectodomain selectively react with the monomeric form of secreted MD-2","volume":"283","author":"Teghanemt","year":"2008","journal-title":"J. Biol. Chem."},{"key":"2023072602431749600_bib18","doi-asserted-by":"publisher","first-page":"1135","DOI":"10.1084\/jem.20042614","article-title":"The nuclear factor HMGB1 mediates hepatic injury after murine liver ischemia-reperfusion","volume":"201","author":"Tsung","year":"2005","journal-title":"J. Exp. Med."},{"key":"2023072602431749600_bib19","doi-asserted-by":"publisher","first-page":"492","DOI":"10.1097\/SHK.0000000000000050","article-title":"High-mobility group box 1 mediates persistent splenocyte priming in sepsis survivors: evidence from a murine model","volume":"40","author":"Vald\u00e9s-Ferrer","year":"2013","journal-title":"Shock."},{"key":"2023072602431749600_bib20","doi-asserted-by":"publisher","first-page":"1519","DOI":"10.1084\/jem.20120189","article-title":"Mutually exclusive redox forms of HMGB1 promote cell recruitment or proinflammatory cytokine release","volume":"209","author":"Venereau","year":"2012","journal-title":"J. Exp. Med."},{"key":"2023072602431749600_bib21","doi-asserted-by":"publisher","first-page":"437","DOI":"10.1016\/j.imbio.2006.05.010","article-title":"MD-2","volume":"211","author":"Visintin","year":"2006","journal-title":"Immunobiology."},{"key":"2023072602431749600_bib22","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1126\/science.285.5425.248","article-title":"HMG-1 as a late mediator of endotoxin lethality in mice","volume":"285","author":"Wang","year":"1999","journal-title":"Science."},{"key":"2023072602431749600_bib23","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1517\/14728222.2014.863876","article-title":"Targeting HMGB1 in the treatment of sepsis","volume":"18","author":"Wang","year":"2014","journal-title":"Expert Opin. Ther. Targets."},{"key":"2023072602431749600_bib24","doi-asserted-by":"publisher","first-page":"296","DOI":"10.1073\/pnas.2434651100","article-title":"Reversing established sepsis with antagonists of endogenous high-mobility group box 1","volume":"101","author":"Yang","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072602431749600_bib25","doi-asserted-by":"publisher","first-page":"11942","DOI":"10.1073\/pnas.1003893107","article-title":"A critical cysteine is required for HMGB1 binding to Toll-like receptor 4 and activation of macrophage cytokine release","volume":"107","author":"Yang","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072602431749600_bib26","doi-asserted-by":"publisher","first-page":"250","DOI":"10.2119\/molmed.2011.00389","article-title":"Redox modification of cysteine residues regulates the cytokine activity of high mobility group box-1 (HMGB1)","volume":"18","author":"Yang","year":"2012","journal-title":"Mol. Med."},{"key":"2023072602431749600_bib27","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1189\/jlb.1212662","article-title":"The many faces of HMGB1: molecular structure-functional activity in inflammation, apoptosis, and chemotaxis","volume":"93","author":"Yang","year":"2013","journal-title":"J. Leukoc. Biol."},{"key":"2023072602431749600_bib28","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1080\/08927022.2011.649428","article-title":"Insights into binding modes of tumstatin peptide T7 with the active site of \u03b1v\u03b23 integrin","volume":"38","author":"Zan","year":"2012","journal-title":"Mol. Simul."},{"key":"2023072602431749600_bib29","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1038\/nature08780","article-title":"Circulating mitochondrial DAMPs cause inflammatory responses to injury","volume":"464","author":"Zhang","year":"2010","journal-title":"Nature."}],"container-title":["Journal of Experimental Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/212\/1\/5\/1752438\/jem_20141318.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jem\/article-pdf\/212\/1\/5\/1752438\/jem_20141318.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T22:43:59Z","timestamp":1690325039000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jem\/article\/212\/1\/5\/41647\/MD-2-is-required-for-disulfide-HMGB1-dependent"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,1,5]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2015,1,12]]}},"URL":"https:\/\/doi.org\/10.1084\/jem.20141318","relation":{},"ISSN":["1540-9538","0022-1007"],"issn-type":[{"value":"1540-9538","type":"electronic"},{"value":"0022-1007","type":"print"}],"subject":[],"published":{"date-parts":[[2015,1,5]]}}}