{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T13:22:40Z","timestamp":1790428960472,"version":"4.1.0"},"reference-count":50,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2019,9,1]],"date-time":"2019-09-01T00:00:00Z","timestamp":1567296000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,9,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Heme is a critical danger molecule liberated from hemeproteins in various conditions, including from hemoglobin in hemolytic diseases. Heme may cause thromboinflammatory damage by activating inflammatory and hemostatic pathways, such as complement, the TLRs, coagulation, and platelets. In this study, we explored the effect of single and dual inhibition of complement component C5 and TLR coreceptor CD14 on heme-induced thromboinflammation in an ex vivo human whole blood model. Heme induced a dose-dependent activation of complement via the alternative pathway. Single inhibition of C5 by eculizumab attenuated the release of IL-6, IL-8, TNF, MCP-1, MIP-1\u03b1, IFN-\u03b3, LTB-4, MMP-8 and -9, and IL-1Ra with more than 60% (p &amp;lt; 0.05 for all) reduced the upregulation of CD11b on granulocytes and monocytes by 59 and 40%, respectively (p &amp;lt; 0.05), and attenuated monocytic tissue factor expression by 33% (p &amp;lt; 0.001). Blocking CD14 attenuated IL-6 and TNF by more than 50% (p &amp;lt; 0.05). In contrast to single inhibition, combined C5 and CD14 was required for a significantly attenuated prothrombin cleavage (72%, p &amp;lt; 0.05). Markers of thromboinflammation were also quantified in two patients admitted to the hospital with sickle cell disease (SCD) crisis. Both SCD patients had pronounced hemolysis and depleted plasma hemopexin and haptoglobin. Plasma heme and complement activation was markedly increased in one patient, a coinciding observation as demonstrated ex vivo. In conclusion, heme-induced thromboinflammation was largely attenuated by C5 inhibition alone, with a beneficial effect of adding a CD14 inhibitor to attenuate prothrombin activation. Targeting C5 has the potential to reduce thromboinflammation in SCD crisis patients.<\/jats:p>","DOI":"10.4049\/jimmunol.1900047","type":"journal-article","created":{"date-parts":[[2019,8,14]],"date-time":"2019-08-14T14:29:03Z","timestamp":1565792943000},"page":"1571-1578","source":"Crossref","is-referenced-by-count":40,"title":["Complement Component C5 and TLR Molecule CD14 Mediate Heme-Induced Thromboinflammation in Human Blood"],"prefix":"10.1093","volume":"203","author":[{"given":"Anub M","family":"Thomas","sequence":"first","affiliation":[{"name":"Department of Immunology, Oslo University Hospital and K.G. Jebsen Inflammation Research Centre, University of Oslo , 0318 Oslo ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6194-6903","authenticated-orcid":false,"given":"Alexandra","family":"Gerogianni","sequence":"additional","affiliation":[{"name":"Linnaeus Centre for Biomaterials Chemistry, Linnaeus University , 391 82 Kalmar ,","place":["Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin B","family":"McAdam","sequence":"additional","affiliation":[{"name":"Department of Immunology, Oslo University Hospital and K.G. Jebsen Inflammation Research Centre, University of Oslo , 0318 Oslo ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yngvar","family":"Fl\u00f8isand","sequence":"additional","affiliation":[{"name":"Department of Haematology, Oslo University Hospital , Rikshospitalet, 0424 Oslo ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1750-8065","authenticated-orcid":false,"given":"Corinna","family":"Lau","sequence":"additional","affiliation":[{"name":"Research Laboratory, Nordland Hospital , 8092 Bodo ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0354-5068","authenticated-orcid":false,"given":"Terje","family":"Espevik","sequence":"additional","affiliation":[{"name":"Centre of Molecular Inflammation Research, Norwegian University of Science and Technology , 7491 Trondheim ,","place":["Norway"]},{"name":"Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology , 7491 Trondheim ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Per H","family":"Nilsson","sequence":"additional","affiliation":[{"name":"Department of Immunology, Oslo University Hospital and K.G. Jebsen Inflammation Research Centre, University of Oslo , 0318 Oslo ,","place":["Norway"]},{"name":"Linnaeus Centre for Biomaterials Chemistry, Linnaeus University , 391 82 Kalmar ,","place":["Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5785-802X","authenticated-orcid":false,"given":"Tom Eirik","family":"Mollnes","sequence":"additional","affiliation":[{"name":"Department of Immunology, Oslo University Hospital and K.G. Jebsen Inflammation Research Centre, University of Oslo , 0318 Oslo ,","place":["Norway"]},{"name":"Research Laboratory, Nordland Hospital , 8092 Bodo ,","place":["Norway"]},{"name":"K.G. Jebsen Thrombosis Research and Expertise Centre, University of Tromso , 9037 Tromso ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5408-7162","authenticated-orcid":false,"given":"Andreas","family":"Barratt-Due","sequence":"additional","affiliation":[{"name":"Department of Immunology, Oslo University Hospital and K.G. Jebsen Inflammation Research Centre, University of Oslo , 0318 Oslo ,","place":["Norway"]},{"name":"Division of Emergencies and Critical Care, Oslo University Hospital , Rikshospitalet, 0027 Oslo ,","place":["Norway"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2019,9,15]]},"reference":[{"key":"2025030417541968300_r1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.3109\/03602539308993973","article-title":"Chemistry and biology of heme. Effect of metal salts, organometals, and metalloporphyrins on heme synthesis and catabolism, with special reference to clinical implications and interactions with cytochrome P-450","volume":"25","author":"Beri","year":"1993","journal-title":"Drug Metab. Rev."},{"key":"2025030417541968300_r2","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1124\/pr.55.3.5","article-title":"Different faces of the heme-heme oxygenase system in inflammation","volume":"55","author":"Wagener","year":"2003","journal-title":"Pharmacol. Rev."},{"key":"2025030417541968300_r3","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.4049\/jimmunol.173.3.2023","article-title":"Heme inhibits human neutrophil apoptosis: involvement of phosphoinositide 3-kinase, MAPK, and NF-kappaB","volume":"173","author":"Arruda","year":"2004","journal-title":"J. Immunol."},{"key":"2025030417541968300_r4","doi-asserted-by":"crossref","first-page":"1383","DOI":"10.1038\/nm1202-799","article-title":"Cell-free hemoglobin limits nitric oxide bioavailability in sickle-cell disease","volume":"8","author":"Reiter","year":"2002","journal-title":"Nat. Med."},{"key":"2025030417541968300_r5","doi-asserted-by":"crossref","first-page":"3805","DOI":"10.1182\/blood-2014-07-589283","article-title":"Circulating cell membrane microparticles transfer heme to endothelial cells and trigger vasoocclusions in sickle cell disease","volume":"125","author":"Camus","year":"2015","journal-title":"Blood"},{"key":"2025030417541968300_r6","doi-asserted-by":"crossref","first-page":"1835","DOI":"10.1093\/infdis\/jiv283","article-title":"Complement activation correlates with disease severity and contributes to cytokine responses in Plasmodium falciparum malaria","volume":"212","author":"Berg","year":"2015","journal-title":"J. Infect. Dis."},{"key":"2025030417541968300_r7","doi-asserted-by":"crossref","first-page":"4160","DOI":"10.1182\/blood.V99.11.4160","article-title":"Neutrophil activation by heme: implications for inflammatory processes","volume":"99","author":"Gra\u00e7a-Souza","year":"2002","journal-title":"Blood"},{"key":"2025030417541968300_r8","doi-asserted-by":"crossref","first-page":"3818","DOI":"10.1182\/blood-2013-10-529982","article-title":"Heme-induced neutrophil extracellular traps contribute to the pathogenesis of sickle cell disease","volume":"123","author":"Chen","year":"2014","journal-title":"Blood"},{"key":"2025030417541968300_r9","doi-asserted-by":"crossref","first-page":"96910","DOI":"10.1172\/jci.insight.96910","article-title":"Intravascular hemolysis activates complement via cell-free heme and heme-loaded microvesicles","volume":"3","author":"Merle","year":"2018","journal-title":"JCI Insight"},{"key":"2025030417541968300_r10","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1182\/blood-2013-03-489245","article-title":"Complement activation by heme as a secondary hit for atypical hemolytic uremic syndrome","volume":"122","author":"Frimat","year":"2013","journal-title":"Blood"},{"key":"2025030417541968300_r11","doi-asserted-by":"crossref","first-page":"919","DOI":"10.3324\/haematol.2018.198622","article-title":"Factor H interferes with the adhesion of sickle red cells to vascular endothelium: a novel disease-modulating molecule","volume":"104","author":"Lombardi","year":"2019","journal-title":"Haematologica"},{"key":"2025030417541968300_r12","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1182\/blood-2013-04-495887","article-title":"Heme triggers TLR4 signaling leading to endothelial cell activation and vaso-occlusion in murine sickle cell disease","volume":"123","author":"Belcher","year":"2014","journal-title":"Blood"},{"key":"2025030417541968300_r13","doi-asserted-by":"crossref","first-page":"20221","DOI":"10.1074\/jbc.M610737200","article-title":"Characterization of heme as activator of Toll-like receptor 4","volume":"282","author":"Figueiredo","year":"2007","journal-title":"J. Biol. Chem."},{"key":"2025030417541968300_r14","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.molmed.2016.01.004","article-title":"Heme: modulator of plasma systems in hemolytic diseases","volume":"22","author":"Roumenina","year":"2016","journal-title":"Trends Mol. Med."},{"key":"2025030417541968300_r15","doi-asserted-by":"crossref","first-page":"3008","DOI":"10.3389\/fimmu.2018.03008","article-title":"Heme drives susceptibility of glomerular endothelium to complement overactivation due to inefficient upregulation of heme oxygenase-1","volume":"9","author":"May","year":"2018","journal-title":"Front. Immunol."},{"key":"2025030417541968300_r16","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1385\/IR:34:2:157","article-title":"The role of complement in danger sensing and transmission","volume":"34","author":"K\u00f6hl","year":"2006","journal-title":"Immunol. Res."},{"key":"2025030417541968300_r17","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1126\/science.1071059","article-title":"The danger model: a renewed sense of self","volume":"296","author":"Matzinger","year":"2002","journal-title":"Science"},{"key":"2025030417541968300_r18","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1159\/000317194","article-title":"The harmful role of c5a on innate immunity in sepsis","volume":"2","author":"Ward","year":"2010","journal-title":"J. Innate Immun."},{"key":"2025030417541968300_r19","first-page":"4794","article-title":"A novel C5a receptor-tissue factor cross-talk in neutrophils links innate immunity to coagulation pathways","volume-title":"J. Immunol.","author":"Ritis","year":"2006"},{"key":"2025030417541968300_r20","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1038\/ni.1923","article-title":"Complement: a key system for immune surveillance and homeostasis","volume":"11","author":"Ricklin","year":"2010","journal-title":"Nat. Immunol."},{"key":"2025030417541968300_r21","doi-asserted-by":"crossref","first-page":"1058","DOI":"10.1056\/NEJM200104053441406","article-title":"Complement. First of two parts","volume":"344","author":"Walport","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"2025030417541968300_r22","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/S1471-4906(01)02129-9","article-title":"Complement in inflammatory tissue damage and disease","volume":"23","author":"Mollnes","year":"2002","journal-title":"Trends Immunol."},{"key":"2025030417541968300_r23","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/j.addr.2011.05.010","article-title":"Advances in assay of complement function and activation","volume":"63","author":"Harboe","year":"2011","journal-title":"Adv. Drug Deliv. Rev."},{"key":"2025030417541968300_r24","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1038\/ni.1863","article-title":"The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors","volume":"11","author":"Kawai","year":"2010","journal-title":"Nat. Immunol."},{"key":"2025030417541968300_r25","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1038\/nri3151","article-title":"Accessory molecules for Toll-like receptors and their function","volume":"12","author":"Lee","year":"2012","journal-title":"Nat. Rev. Immunol."},{"key":"2025030417541968300_r26","first-page":"819","article-title":"Combined inhibition of complement C5 and CD14 markedly attenuates inflammation, thrombogenicity, and hemodynamic changes in porcine sepsis","volume-title":"J. Immunol.","author":"Barratt-Due","year":"2013"},{"key":"2025030417541968300_r27","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1189\/jlb.3VMR0316-132R","article-title":"Dual inhibition of complement and Toll-like receptors as a novel approach to treat inflammatory diseases-C3 or C5 emerge together with CD14 as promising targets","volume":"101","author":"Barratt-Due","year":"2017","journal-title":"J. Leukoc. Biol."},{"key":"2025030417541968300_r28","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1159\/000486542","article-title":"Combined inhibition of C5 and CD14 attenuates systemic inflammation in a piglet model of meconium aspiration syndrome","volume":"113","author":"Thomas","year":"2018","journal-title":"Neonatology"},{"key":"2025030417541968300_r29","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1111\/j.1365-2044.2011.07011.x","article-title":"Complement takes its Toll: an inflammatory crosstalk between Toll-like receptors and the receptors for the complement anaphylatoxin C5a","volume":"67","author":"Holst","year":"2012","journal-title":"Anaesthesia"},{"key":"2025030417541968300_r30","first-page":"4769","article-title":"Chimeric anti-CD14 IGG2\/4 hybrid antibodies for therapeutic intervention in pig and human models of inflammation","volume-title":"J. Immunol.","author":"Lau","year":"2013"},{"key":"2025030417541968300_r31","first-page":"1869","article-title":"Essential role of the C5a receptor in E coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. [Published erratum appears in 2002 Blood 100: 2691.]","volume":"100","author":"Mollnes","year":"2002","journal-title":"Blood"},{"key":"2025030417541968300_r32","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1111\/j.1365-3083.1985.tb01871.x","article-title":"Quantification of the terminal complement complex in human plasma by an enzyme-linked immunosorbent assay based on monoclonal antibodies against a neoantigen of the complex","volume":"22","author":"Mollnes","year":"1985","journal-title":"Scand. J. Immunol."},{"key":"2025030417541968300_r33","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.molimm.2013.05.221","article-title":"An international serum standard for application in assays to detect human complement activation products. [Published erratum appears in 2014 Mol. Immunol. 60: 115.]","volume":"56","author":"Bergseth","year":"2013","journal-title":"Mol. Immunol."},{"key":"2025030417541968300_r34","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1111\/j.1365-3083.1988.tb02353.x","article-title":"Characterization of a monoclonal antibody MoAb bH6 reacting with a neoepitope of human C3 expressed on C3b, iC3b, and C3c","volume":"27","author":"Garred","year":"1988","journal-title":"Scand. J. Immunol."},{"key":"2025030417541968300_r35","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1247\/csf.26.235","article-title":"Hemopexins suppress phorbol ester-induced necrosis of polymorphonuclear leucocytes","volume":"26","author":"Suzuki","year":"2001","journal-title":"Cell Struct. Funct."},{"key":"2025030417541968300_r36","doi-asserted-by":"crossref","first-page":"5543","DOI":"10.4049\/jimmunol.179.8.5543","article-title":"Hematin promotes complement alternative pathway-mediated deposition of C3 activation fragments on human erythrocytes: potential implications for the pathogenesis of anemia in malaria","volume":"179","author":"Pawluczkowycz","year":"2007","journal-title":"J. Immunol."},{"key":"2025030417541968300_r37","doi-asserted-by":"crossref","first-page":"16459","DOI":"10.1074\/jbc.M110.206136","article-title":"Heme interacts with c1q and inhibits the classical complement pathway","volume":"286","author":"Roumenina","year":"2011","journal-title":"J. Biol. Chem."},{"key":"2025030417541968300_r38","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1006\/clin.1995.1131","article-title":"Complement activation during painful crisis in sickle cell anemia","volume":"76","author":"Mold","year":"1995","journal-title":"Clin. Immunol. Immunopathol."},{"key":"2025030417541968300_r39","first-page":"116","article-title":"Complement activation during intravascular hemolysis: implication for sickle cell disease and hemolytic transfusion reactions","volume-title":"Transfus. Clin. Biol.","author":"Merle","year":"2019"},{"key":"2025030417541968300_r40","first-page":"6562","article-title":"Leukotriene B4 mediates neutrophil migration induced by heme","volume-title":"J. Immunol.","author":"Monteiro","year":"2011"},{"key":"2025030417541968300_r41","first-page":"4913","article-title":"Ornithodoros moubata complement inhibitor is an equally effective C5 inhibitor in pigs and humans","volume-title":"J. Immunol.","author":"Barratt-Due","year":"2011"},{"key":"2025030417541968300_r42","doi-asserted-by":"crossref","first-page":"1939","DOI":"10.1038\/jcbfm.2010.45","article-title":"Hemoglobin-induced oxidative stress contributes to matrix metalloproteinase activation and blood-brain barrier dysfunction in vivo","volume":"30","author":"Katsu","year":"2010","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"2025030417541968300_r43","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1016\/j.ajpath.2011.04.024","article-title":"Complement activation triggers metalloproteinases release inducing cervical remodeling and preterm birth in mice","volume":"179","author":"Gonzalez","year":"2011","journal-title":"Am. J. Pathol."},{"key":"2025030417541968300_r44","doi-asserted-by":"crossref","first-page":"339","DOI":"10.3109\/10409238809082548","article-title":"Initiation of coagulation by tissue factor","volume":"23","author":"Bach","year":"1988","journal-title":"CRC Crit. Rev. Biochem."},{"key":"2025030417541968300_r45","first-page":"279","article-title":"Elevated levels of prothrombin fragment 1 + 2 indicate high risk of thrombosis","volume-title":"Clin. Appl. Thromb. Hemost.","author":"Ota","year":"2008"},{"key":"2025030417541968300_r46","doi-asserted-by":"crossref","first-page":"308","DOI":"10.3324\/haematol.2014.114728","article-title":"Excess of heme induces tissue factor-dependent activation of coagulation in mice","volume":"100","author":"Sparkenbaugh","year":"2015","journal-title":"Haematologica"},{"key":"2025030417541968300_r47","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1182\/blood-2012-04-424143","article-title":"Tissue factor promotes activation of coagulation and inflammation in a mouse model of sickle cell disease","volume":"120","author":"Chantrathammachart","year":"2012","journal-title":"Blood"},{"key":"2025030417541968300_r48","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1002\/ajh.25384","article-title":"Critical role of C5a in sickle cell disease","volume":"94","author":"Vercellotti","year":"2019","journal-title":"Am. J. Hematol."},{"key":"2025030417541968300_r49","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.it.2007.02.006","article-title":"Complement and coagulation: strangers or partners in crime?","volume":"28","author":"Markiewski","year":"2007","journal-title":"Trends Immunol."},{"key":"2025030417541968300_r50","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/978-1-4614-4118-2_8","article-title":"The effects of selective complement and CD14 inhibition on the E. coli-induced tissue factor mRNA upregulation, monocyte tissue factor expression, and tissue factor functional activity in human whole blood","volume":"735","author":"Brekke","year":"2013","journal-title":"Adv. Exp. Med. Biol."}],"container-title":["The Journal of Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/203\/6\/1571\/61418228\/ji1900047.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jimmunol\/article-pdf\/203\/6\/1571\/61418228\/ji1900047.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T15:03:10Z","timestamp":1741100590000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jimmunol\/article\/203\/6\/1571\/7952207"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9]]},"references-count":50,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2019,9,15]]}},"URL":"https:\/\/doi.org\/10.4049\/jimmunol.1900047","relation":{},"ISSN":["0022-1767","1550-6606"],"issn-type":[{"value":"0022-1767","type":"print"},{"value":"1550-6606","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2019,9]]},"published":{"date-parts":[[2019,9]]}}}