{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T01:26:49Z","timestamp":1771982809193,"version":"3.50.1"},"reference-count":25,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2016,1,16]],"date-time":"2016-01-16T00:00:00Z","timestamp":1452902400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Xenotransplantation"],"published-print":{"date-parts":[[2016,3]]},"abstract":"Abstract<\/jats:title>Background<\/jats:title>Xenogeneic thrombotic microangiopathy (TMA<\/jats:styled-content>) and acute vascular rejection (AVR<\/jats:styled-content>) prevent long\u2010term survival of porcine xenografts after transplantation into non\u2010human primates. Preformed xenoreactive natural antibodies (XNA<\/jats:styled-content>) cause endothelial damage and activate the complement system. Mechanisms of xenogeneic coagulation and platelet activation are only poorly characterized.<\/jats:p><\/jats:sec>Methods<\/jats:title>A microfluidic flow chamber was used to study platelet activation and thrombus formation of human platelet\u2010rich plasma (PRP<\/jats:styled-content>) upon perfusion over wild\u2010type (WT<\/jats:styled-content>) or \u03b1\u20101,3\u2010 galactosyltransferase knockout (GTKO<\/jats:styled-content>) and human CD<\/jats:styled-content>46 (hCD<\/jats:styled-content>46) transgenic porcine aortic endothelial cells (PAEC<\/jats:styled-content>). Activation of plasma coagulation (thrombin\u2013anti\u2010thrombin complex; TAT<\/jats:styled-content>) and complement (C3a, C5a) was studied in human platelet\u2010free plasma (PFP<\/jats:styled-content>) after co\u2010incubation with PAEC<\/jats:styled-content>. The activation of PAEC<\/jats:styled-content> (E\u2010Selectin, tissue factor, ICAM<\/jats:styled-content>\u20101, ICAM<\/jats:styled-content>\u20102, VCAM<\/jats:styled-content>\u20101) was studied after incubation with human serum. Eculizumab (200 \u03bcg\/ml) was used to inhibit terminal complement activation in all experiments.<\/jats:p><\/jats:sec>Results<\/jats:title>WT<\/jats:styled-content>\u2010PAEC<\/jats:styled-content> perfused with human PRP<\/jats:styled-content> showed thrombus formation at different shear rates (3 dyn\/cm2<\/jats:sup>: 23 \u00b1 10%; 10 dyn\/cm2<\/jats:sup>: 17 \u00b1 10% of flow chamber viewing field). GTKO<\/jats:styled-content>\/hCD<\/jats:styled-content>46 PAEC<\/jats:styled-content> exhibited reduced, but not fully prevented thrombus formation (3 dyn\/cm2<\/jats:sup>: 12 \u00b1 12%). Porcine PRP<\/jats:styled-content> caused little or no thrombus formation (3.0 \u00b1 4% and 0.5 \u00b1 0.9%, respectively). Flow cytometry of human platelets after perfusion over WT<\/jats:styled-content>\u2010PAEC<\/jats:styled-content> revealed an increase in platelet CD<\/jats:styled-content>62P expression (29.5 \u00b1 3%), compared to non\u2010perfused PRP<\/jats:styled-content> (7 \u00b1 2%) or PRP<\/jats:styled-content> running through empty flow chambers (12.7 \u00b1 0.3%). Incubation of human PFP<\/jats:styled-content> with PAEC<\/jats:styled-content> resulted in an increase of TAT<\/jats:styled-content> that correlated with C5a activation. Specific inhibition of complement by eculizumab prevented thrombus formation (WT<\/jats:styled-content>\u2010PAEC<\/jats:styled-content>: 1.6 \u00b1 2% at 3 dyn\/cm2<\/jats:sup> and 0.24 \u00b1 0.33% at 10 dyn\/cm2<\/jats:sup>, GTKO<\/jats:styled-content>\/hCD<\/jats:styled-content>46 PAEC<\/jats:styled-content>: 0.2 \u00b1 0.3% at 3 dyn\/cm2<\/jats:sup>) as well as activation of coagulation and platelets. Induction of endothelial E\u2010Selectin and VCAM<\/jats:styled-content>\u20101 in WT<\/jats:styled-content>\u2010PAEC<\/jats:styled-content> upon incubation with human serum was significantly reduced by eculizumab. Eculizumab did not reduce thrombin generation capacity of human PRP<\/jats:styled-content> or normal platelet aggregation.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>Thrombus formation in this ex vivo model of xenogeneic TMA<\/jats:styled-content> was closely linked with complement activation. Specific inhibition of complement C5 by eculizumab prevented endothelial cell activation, but also coagulation and platelet activation without compromising thrombin generation capacity of human blood or normal platelet function.<\/jats:p><\/jats:sec>","DOI":"10.1111\/xen.12218","type":"journal-article","created":{"date-parts":[[2016,1,16]],"date-time":"2016-01-16T22:04:01Z","timestamp":1452981841000},"page":"117-127","source":"Crossref","is-referenced-by-count":12,"title":["Inhibition of complement component C5 prevents clotting in an ex vivo model of xenogeneic activation of coagulation"],"prefix":"10.1111","volume":"23","author":[{"given":"Dennis","family":"Rataj","sequence":"first","affiliation":[{"name":"Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation Hannover Medical School Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonja","family":"Werwitzke","sequence":"additional","affiliation":[{"name":"Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation Hannover Medical School Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Birgitt","family":"Haarmeijer","sequence":"additional","affiliation":[{"name":"Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation Hannover Medical School Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Winkler","sequence":"additional","affiliation":[{"name":"Department for General, Visceral and Transplantation Surgery Hannover Medical School Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wolf","family":"Ramackers","sequence":"additional","affiliation":[{"name":"Department for General, Visceral and Transplantation Surgery Hannover Medical School Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bj\u00f6rn","family":"Petersen","sequence":"additional","affiliation":[{"name":"Institute of Farm Animal Genetics Friedrich\u2010Loeffler\u2010Institute Neustadt Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heiner","family":"Niemann","sequence":"additional","affiliation":[{"name":"Institute of Farm Animal Genetics Friedrich\u2010Loeffler\u2010Institute Neustadt Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annegret","family":"W\u00fcnsch","sequence":"additional","affiliation":[{"name":"Molecular Animal Breeding and Biotechnology Gene Center and Department of Veterinary Sciences Ludwig Maximilian University of Munich Munich Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea","family":"B\u00e4hr","sequence":"additional","affiliation":[{"name":"Molecular Animal Breeding and Biotechnology Gene Center and Department of Veterinary Sciences Ludwig Maximilian University of Munich Munich Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolai","family":"Klymiuk","sequence":"additional","affiliation":[{"name":"Molecular Animal Breeding and Biotechnology Gene Center and Department of Veterinary Sciences Ludwig Maximilian University of Munich Munich Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eckhard","family":"Wolf","sequence":"additional","affiliation":[{"name":"Molecular Animal Breeding and Biotechnology Gene Center and Department of Veterinary Sciences Ludwig Maximilian University of Munich Munich Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan\u2010Michael","family":"Abicht","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology Ludwig Maximilian University of Munich Munich Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Ayares","sequence":"additional","affiliation":[{"name":"Revivicor, Inc. 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