{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:00:07Z","timestamp":1792789207299,"version":"4.5.0"},"reference-count":38,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Science Fund of the Republic of Serbia","award":["7749695"],"award-info":[{"award-number":["7749695"]}],"id":[{"id":"https:\/\/ror.org\/02r531a15","id-type":"ROR","asserted-by":"crossref"}]},{"award":["7749695"],"award-info":[{"award-number":["7749695"]}],"id":[{"id":"https:\/\/ror.org\/02r531a15","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Graft-versus-host disease (GvHD) remains a major complication in patients with hematological malignancies undergoing hematopoietic stem cell transplantation, driven by interconnected neutrophil activation, endothelial dysfunction, inflammation, and dysregulated coagulation. However, characterization of these pathways in GvHD patients remains limited. We used ELISA and flow cytometry to quantify biomarkers of neutrophil extracellular traps (NETs), cytokines, endothelial activation, fibrinolysis, coagulation, and immune cell subsets in peripheral blood of GvHD patients, and Western blot to assess NF-\u03baB, STAT3, and p38 MAPK signaling. GvHD patients showed a thromboinflammatory profile, with elevated NET markers (MPO, cell-free DNA), increased cytokines (IL-1\u03b2, IL-8, MCP-1, TNF-\u03b1), and amplified platelet activation (P-selectin). Coagulation and fibrinolytic activity were enhanced, reflected by increased tissue factor (TF) and urokinase plasminogen activator (uPA), alongside higher frequencies of TF-expressing monocytes and platelet\u2013monocyte aggregates. In vitro, TNF-\u03b1 activated NF-\u03baB, STAT3, and p38 MAPK signaling and increased ICAM-1- and VCAM-1-positive mononuclear cells, indicating a proinflammatory, adhesive phenotype, while transendothelial migration and nitric oxide levels were reduced. Leukemia patients with GvHD thus show a coordinated thromboinflammatory response involving NET activation, platelet\u2013monocyte interactions, inflammatory signaling, endothelial dysfunction, and coagulation\u2013fibrinolytic dysregulation, providing mechanistic insight into GvHD pathophysiology and highlighting circulating biomarkers as candidates for early detection, monitoring, and therapeutic intervention.<\/jats:p>","DOI":"10.3390\/cells15181690","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T09:01:38Z","timestamp":1789981298000},"page":"1690","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Activation of Inflammatory and Coagulation Pathways in Patients with Graft-Versus-Host Disease"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6305-8205","authenticated-orcid":false,"given":"Emilija","family":"\u017divkovi\u0107","sequence":"first","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0958-8811","authenticated-orcid":false,"given":"Milena","family":"Todorovi\u0107 Balint","sequence":"additional","affiliation":[{"name":"Clinic for Hematology, University Clinical Center of Serbia, Dr Koste Todorovi\u0107a 2, 11000 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, Dr Suboti\u0107a starijeg 8, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5132-8319","authenticated-orcid":false,"given":"Tijana","family":"Suboti\u010dki","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1171-5186","authenticated-orcid":false,"given":"Milo\u0161","family":"Dikli\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"}]},{"given":"Dragoslava","family":"\u0110iki\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0232-5322","authenticated-orcid":false,"given":"Milica","family":"Vukoti\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Nevena","family":"Be\u0161evi\u0107","sequence":"additional","affiliation":[{"name":"Clinic for Hematology, University Clinical Center of Serbia, Dr Koste Todorovi\u0107a 2, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6246-0198","authenticated-orcid":false,"given":"Andrej","family":"Pe\u0161i\u0107","sequence":"additional","affiliation":[{"name":"Clinic for Hematology, University Clinical Center of Serbia, Dr Koste Todorovi\u0107a 2, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0362-1449","authenticated-orcid":false,"given":"Vladan P.","family":"\u010coki\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3977-2780","authenticated-orcid":false,"given":"Olivera","family":"Mitrovi\u0107 Ajti\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotica 4, 11129 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.iac.2009.10.001","article-title":"Pathogenesis and Management of Graft versus Host Disease","volume":"30","author":"Choi","year":"2010","journal-title":"Immunol. Allergy Clin. N. Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1002\/rth2.12266","article-title":"The dual role of platelet-innate immune cell interactions in thrombo-inflammation","volume":"4","author":"Rayes","year":"2019","journal-title":"Res. Pract. Thromb. Haemost."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1186\/s43556-025-00337-9","article-title":"Neutrophil Extracellular Traps (NETs) in health and disease","volume":"6","author":"Shahzad","year":"2025","journal-title":"Mol. Biomed."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1901","DOI":"10.1161\/ATVBAHA.118.311150","article-title":"Neutrophil Extracellular Traps Induce Endothelial Cell Activation and Tissue Factor Production Through Interleukin-1\u03b1 and Cathepsin G","volume":"38","author":"Folco","year":"2018","journal-title":"Arter. Thromb. Vasc. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1038\/s41577-022-00787-0","article-title":"Neutrophil extracellular traps in systemic autoimmune and autoinflammatory diseases","volume":"23","author":"Wigerblad","year":"2022","journal-title":"Nat. Rev. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"10760296241304771","DOI":"10.1177\/10760296241304771","article-title":"Graft-Versus-Host Disease Sustains Coagulation Activity for two Years After Pediatric Allogeneic Hematopoietic Stem Cell Transplantation","volume":"31","author":"Koskenvuo","year":"2025","journal-title":"Clin. Appl. Thromb."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Almskog, L.M., and \u00c5gren, A. (2025). Thromboinflammation vs. immunothrombosis: Strategies for overcoming anticoagulant resistance in COVID-19 and other hyperinflammatory diseases. Is ROTEM helpful or not?. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1599639"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1161\/01.ATV.18.5.842","article-title":"Upregulation of VCAM-1 and ICAM-1 at Atherosclerosis-Prone Sites on the Endothelium in the ApoE-Deficient Mouse","volume":"18","author":"Nakashima","year":"1998","journal-title":"Arter. Thromb. Vasc. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Neidemire-Colley, L., Robert, J., Ackaoui, A., Dorrance, A.M., Guimond, M., and Ranganathan, P. (2022). Role of Endothelial Cells in Graft-versus-Host Disease. Front. Immunol., 13, Erratum in Front. Immunol.\u00a02023, 13, 1125035. https:\/\/doi.org\/10.3389\/fimmu.2022.1125035.","DOI":"10.3389\/fimmu.2022.1033490"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2147","DOI":"10.3324\/haematol.2020.253716","article-title":"Endothelial damage and dysfunction in acute graft-versus-host disease","volume":"106","author":"Cordes","year":"2020","journal-title":"Haematologica"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.gendis.2023.02.039","article-title":"HIF-1\u03b1 signaling: Essential roles in tumorigenesis and implications in targeted therapies","volume":"11","author":"Zhao","year":"2024","journal-title":"Genes Dis."},{"key":"ref_12","first-page":"105","article-title":"Hypoxia-Inducible Factors as Essential Regulators of Inflammation","volume":"345","author":"Imtiyaz","year":"2010","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1038\/s41423-025-01310-w","article-title":"NF-\u03baB in Inflammation and Cancer","volume":"22","author":"Mao","year":"2025","journal-title":"Cell. Mol. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1182\/blood-2005-11-4454","article-title":"Hydroxyurea induces the eNOS-cGMP pathway in endothelial cells","volume":"108","author":"Cokic","year":"2006","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3037744","DOI":"10.1155\/jimr\/3037744","article-title":"Low-Density Neutrophils: Enigmatic Cells in Health and Disease","volume":"2025","author":"Rosales","year":"2025","journal-title":"J. Immunol. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e2250010","DOI":"10.1002\/eji.202250010","article-title":"Neutrophil activation and neutrophil extracellular traps (NETs) in COVID-19 ARDS and immunothrombosis","volume":"53","author":"Cesta","year":"2022","journal-title":"Eur. J. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Kimball, A.S., Obi, A.T., Diaz, J.A., and Henke, P.K. (2016). The Emerging Role of NETs in Venous Thrombosis and Immunothrombosis. Front. Immunol., 7.","DOI":"10.3389\/fimmu.2016.00236"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Zhou, Y., Tao, W., Shen, F., Du, W., Xu, Z., and Liu, Z. (2021). The Emerging Role of Neutrophil Extracellular Traps in Arterial, Venous and Cancer-Associated Thrombosis. Front. Cardiovasc. Med., 8.","DOI":"10.3389\/fcvm.2021.786387"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Wienkamp, A.-K., Erpenbeck, L., and Rossaint, J. (2022). Platelets in the NETworks interweaving inflammation and thrombosis. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.953129"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Li, H., Shan, W., Zhao, X., and Sun, W. (2025). Neutrophils: Linking Inflammation to Thrombosis and Unlocking New Treatment Horizons. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26051965"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.12968\/hmed.2024.0660","article-title":"Relationship Between Neutrophil Extracellular Traps and Venous Thromboembolism: Pathophysiological and Therapeutic Role","volume":"86","author":"Dinc","year":"2025","journal-title":"Br. J. Hosp. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"9204","DOI":"10.1111\/jcmm.15557","article-title":"Monocyte subsets in bone marrow grafts may contribute to a low incidence of acute graft-vs-host disease for young donors","volume":"24","author":"Wen","year":"2020","journal-title":"J. Cell. Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Wilhelm, G., Mertowska, P., Mertowski, S., Przysucha, A., Stru\u017cyna, J., Grywalska, E., and Torres, K. (2023). The Crossroads of the Coagulation System and the Immune System: Interactions and Connections. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241612563"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Mosevoll, K.A., Johansen, S., Wendelbo, \u00d8., Nepstad, I., Bruserud, \u00d8., and Reikvam, H. (2018). Cytokines, Adhesion Molecules, and Matrix Metalloproteases as Predisposing, Diagnostic, and Prognostic Factors in Venous Thrombosis. Front. Med., 5.","DOI":"10.3389\/fmed.2018.00147"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Niu, Y., Liu, C., Li, P., Zhao, J., Jin, J., and Yang, J. (2025). Interleukin Networks in GVHD: Mechanistic Crosstalk, Therapeutic Targeting, and Emerging Paradigms. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26178620"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ztriva, E., Moschonas, I.C., Tselepis, A., Lambrou, D., Ntaios, G., Savopoulos, C., and Kaiafa, G. (2025). Monocyte Chemoattractant Protein-1 as a Biomarker in Acute Ischemic Stroke: A Prospective Pilot Study. J. Clin. Med., 14.","DOI":"10.3390\/jcm14103295"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"100015","DOI":"10.1016\/j.bvth.2024.100015","article-title":"Targeting the P-selectin\/PSGL-1 pathway: Discovery of disease-modifying therapeutics for disorders of thromboinflammation","volume":"1","author":"Escopy","year":"2024","journal-title":"Blood Vessel. Thromb. Hemost."},{"key":"ref_28","unstructured":"Michelson, A.D., Cattaneo, M., Frelinger, A.L., and Newman, P.J. (2019). The Role of Platelets in Inflammation. Platelets, Academic Press. [4th ed.]."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Rolling, C.C., Barrett, T.J., and Berger, J.S. (2023). Platelet-monocyte aggregates: Molecular mediators of thromboinflammation. Front. Cardiovasc. Med., 10.","DOI":"10.3389\/fcvm.2023.960398"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Lampiasi, N., and Russo, R. (2026). HIF-1\u03b1 as a Central Regulator of Monocyte Responses to Hypoxia. Biology, 15.","DOI":"10.3390\/biology15030213"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1016\/j.bbmt.2010.02.008","article-title":"Endothelial Dysfunction after Hematopoietic Stem Cell Transplantation: Role of the Conditioning Regimen and the Type of Transplantation","volume":"16","author":"Palomo","year":"2010","journal-title":"Biol. Blood Marrow Transplant."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Milone, G., Bellofiore, C., Leotta, S., Milone, G.A., Cupri, A., Duminuco, A., Garibaldi, B., and Palumbo, G. (2022). Endothelial Dysfunction after Hematopoietic Stem Cell Transplantation: A Review Based on Physiopathology. J. Clin. Med., 11.","DOI":"10.3390\/jcm11030623"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9480","DOI":"10.3892\/mmr.2017.7825","article-title":"Therapeutic effects of STAT3 inhibition by nifuroxazide on murine acute graft graft-vs.-host disease: Old drug, new use","volume":"16","author":"Jia","year":"2017","journal-title":"Mol. Med. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.cytogfr.2009.11.005","article-title":"Dangerous liaisons: STAT3 and NF-\u03baB collaboration and crosstalk in cancer","volume":"21","author":"Grivennikov","year":"2010","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.1038\/embor.2009.243","article-title":"IKK\/NF-\u03baB and STAT3 pathways: Central signalling hubs in inflammation-mediated tumour promotion and metastasis","volume":"10","author":"Bollrath","year":"2009","journal-title":"Embo Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"20059","DOI":"10.1074\/jbc.R700016200","article-title":"JAK-STAT Signaling: From Interferons to Cytokines","volume":"282","author":"Schindler","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1016\/j.ajpath.2016.12.016","article-title":"Blockade of p38 Mitogen-Activated Protein Kinase Inhibits Murine Sclerodermatous Chronic Graft-versus-Host Disease","volume":"187","author":"Matsushita","year":"2017","journal-title":"Am. J. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Braun, L.M., and Zeiser, R. (2021). Kinase Inhibition as Treatment for Acute and Chronic Graft-Versus-Host Disease. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.760199"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/18\/1690\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T04:26:53Z","timestamp":1790051213000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/18\/1690"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,17]]},"references-count":38,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["cells15181690"],"URL":"https:\/\/doi.org\/10.3390\/cells15181690","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,17]]}}}