{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T22:17:12Z","timestamp":1791497832539,"version":"4.3.4"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T00:00:00Z","timestamp":1778025600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diseases"],"abstract":"<jats:p>Background\/Objectives: Acute intravascular hemolysis is associated with the release of labile heme, which contributes to systemic inflammation and organ dysfunction. Galectin-3 (Gal-3) is a known modulator of inflammatory responses. However, its specific role in heme-induced organ injury remains to be fully elucidated. Methods: We used a phenylhydrazine (PHZ)-induced model of acute hemolysis in wild-type (WT) and Gal-3 knockout (KO) mice to investigate the influence of Gal-3 on tissue alterations and the inflammatory response. Results: Despite equivalent levels of hemolysis and anemia in both genotypes, Gal-3 deficiency was associated with reduced injury in the liver, kidneys, and pancreas. In WT mice, Gal-3 was associated with a pro-inflammatory splenic microenvironment. Conversely, Gal-3 KO mice exhibited a shift toward an immunoregulatory phenotype, characterized by an increased frequency of CD4 + CD25 + FoxP3+ regulatory T cells and IL-10+ macrophages. This shift correlated with preserved organ architecture and a more controlled inflammatory profile. Conclusions: Our findings suggest that Gal-3 may act as a mediator of heme-induced systemic inflammation. By influencing the splenic immune microenvironment and promoting a regulatory phenotype, the absence of Gal-3 appears to alleviate multi-organ stress, suggesting its potential as a target for modulating complications during acute hemolytic crises.<\/jats:p>","DOI":"10.3390\/diseases14050161","type":"journal-article","created":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T09:30:22Z","timestamp":1778059822000},"page":"161","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Galectin-3 Mediates Heme-Induced Multi-Organ Dysfunction by Modulating the Splenic Immune Microenvironment"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-8292-3229","authenticated-orcid":false,"given":"Mirjana","family":"Milinkovic","sequence":"first","affiliation":[{"name":"Institute for Transfusiology and Hemobiology, Military Medical Academy, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6894-1275","authenticated-orcid":false,"given":"Marija","family":"Milovanovic","sequence":"additional","affiliation":[{"name":"Center for Harm Reduction of Biological and Chemical Hazards, Faculty of Medical Sciences, University of Kragujevac, Svetozara Markovi\u0107a 69, 34000 Kragujevac, Serbia"},{"name":"Department of Microbiology and Immunology, Faculty of Medical Sciences, University of Kragujevac, Svetozara Markovi\u0107a 69, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6422-5423","authenticated-orcid":false,"given":"Jelena","family":"Milovanovic","sequence":"additional","affiliation":[{"name":"Center for Harm Reduction of Biological and Chemical Hazards, Faculty of Medical Sciences, University of Kragujevac, Svetozara Markovi\u0107a 69, 34000 Kragujevac, Serbia"},{"name":"Department of Histology and Embryology, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"18011","DOI":"10.1038\/nrdp.2018.10","article-title":"Sickle cell disease","volume":"4","author":"Kato","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1276","DOI":"10.1182\/blood-2012-11-451229","article-title":"Hemolysis and free hemoglobin revisited: Exploring hemoglobin and hemin scavengers as a novel class of therapeutic proteins","volume":"121","author":"Schaer","year":"2013","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/S0140-6736(17)30193-9","article-title":"Sickle cell disease","volume":"390","author":"Ware","year":"2017","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Arsenijevic, A., Milovanovic, J., Stojanovic, B., Djordjevic, D., Stanojevic, I., Jankovic, N., Vojvodic, D., Arsenijevic, N., Lukic, M.L., and Milovanovic, M. (2019). Gal-3 Deficiency Suppresses Novosphyngobium aromaticivorans Inflammasome Activation and IL-17 Driven Autoimmune Cholangitis in Mice. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.01309"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Dutra, F.F., and Bozza, M.T. (2014). Heme on innate immunity and inflammation. Front. Pharmacol., 5.","DOI":"10.3389\/fphar.2014.00115"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1172\/JCI62972","article-title":"Hemolysis and cell-free hemoglobin drive an intrinsic mechanism for human disease","volume":"122","author":"Gladwin","year":"2012","journal-title":"J. Clin. Investig."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.3389\/fimmu.2020.01323","article-title":"Pro-inflammatory Actions of Heme and Other Hemoglobin-Derived DAMPs","volume":"11","author":"Bozza","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"L429","DOI":"10.1152\/ajplung.00018.2021","article-title":"Toxic effects of cell-free hemoglobin on the microvascular endothelium: Implications for pulmonary and nonpulmonary organ dysfunction","volume":"321","author":"Meegan","year":"2021","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_9","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":"ref_10","doi-asserted-by":"crossref","first-page":"E4110","DOI":"10.1073\/pnas.1405023111","article-title":"Hemolysis-induced lethality involves inflammasome activation by heme","volume":"111","author":"Dutra","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","first-page":"471","article-title":"Heme and the Kidney","volume":"101","author":"Nath","year":"2022","journal-title":"Kidney Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1159\/000109196","article-title":"Cytokines as intercellular signals in hemolytic transfusion reactions","volume":"5","author":"Davenport","year":"1996","journal-title":"Biol. Signals"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"23348","DOI":"10.1038\/srep23348","article-title":"Deletion of Galectin-3 Enhances Xenobiotic Induced Murine Primary Biliary Cholangitis by Facilitating Apoptosis of BECs and Release of Autoantigens","volume":"6","author":"Arsenijevic","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"572637","DOI":"10.2147\/JIR.S572637","article-title":"The Role of Galectin-3 in Liver Inflammation and Fibrosis","volume":"19","author":"Lurje","year":"2026","journal-title":"J. Inflamm. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"jcs208884","DOI":"10.1242\/jcs.208884","article-title":"Galectins at a glance","volume":"131","author":"Johannes","year":"2018","journal-title":"J. Cell Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1023\/B:GLYC.0000014072.34840.04","article-title":"Introduction to galectins","volume":"19","author":"Leffler","year":"2004","journal-title":"Glycoconj. J."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Mishra, B.B., Li, Q., Steichen, A.L., Binstock, B.J., Metzger, D.W., Teale, J.M., and Sharma, J. (2013). Galectin-3 functions as an alarmin: Pathogenic role for sepsis development in murine respiratory tularemia. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0059616"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1016\/j.bbagen.2005.12.020","article-title":"Galectin-3: An open-ended story","volume":"1760","author":"Dumic","year":"2006","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Grujcic, M., Milovanovic, M., Nedeljkovic, J., Jovanovic, D., Arsenijevic, D., Solovjova, N., Stankovic, V., Tanaskovic, I., Arsenijevic, A., and Milovanovic, J. (2024). The Possible Effects of Galectin-3 on Mechanisms of Renal and Hepatocellular Injury Induced by Intravascular Hemolysis. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25158129"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1038\/35051594","article-title":"Identification of the hemoglobin scavenger receptor","volume":"409","author":"Kristiansen","year":"2001","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1549S","DOI":"10.1093\/jn\/133.5.1549S","article-title":"Ferritin: At the crossroads of iron and oxygen metabolism","volume":"133","author":"Theil","year":"2003","journal-title":"J. Nutr."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1007\/s00277-023-05589-5","article-title":"Differential expression of adhesion molecules in sickle cell anemia and gut microbiome effect","volume":"103","author":"Delgadinho","year":"2024","journal-title":"Ann. Hematol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2352","DOI":"10.1089\/ars.2012.4834","article-title":"CD163 and inflammation: Biological, diagnostic, and therapeutic aspects","volume":"18","author":"Etzerodt","year":"2013","journal-title":"Antioxid. Redox Signal."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1097\/MOH.0b013e32834521c8","article-title":"Stress erythropoiesis: New signals and new progenitors","volume":"18","author":"Paulson","year":"2011","journal-title":"Curr. Opin. Hematol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1124\/jpet.107.119792","article-title":"Cisplatin-induced acute renal failure is associated with an increase in the cytokines interleukin (IL)-1beta, IL-18, IL-6, and neutrophil infiltration in the kidney","volume":"322","author":"Faubel","year":"2007","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1097\/00007890-199112000-00008","article-title":"The beneficial effect of a prostaglandin I2 analog on ischemic rat liver","volume":"52","author":"Suzuki","year":"1991","journal-title":"Transplantation"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1097\/00000658-199201000-00007","article-title":"A better model of acute pancreatitis for evaluating therapy","volume":"215","author":"Schmidt","year":"1992","journal-title":"Ann. Surg."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1097\/00007890-198303000-00002","article-title":"An experimental model for assessment of renal recovery from warm ischemia","volume":"35","author":"Jablonski","year":"1983","journal-title":"Transplantation"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4458","DOI":"10.4049\/jimmunol.1600709","article-title":"Galectin-3: A Positive Regulator of Leukocyte Recruitment in the Inflamed Microcirculation","volume":"198","author":"Gittens","year":"2017","journal-title":"J. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1970","DOI":"10.1038\/s41418-025-01514-7","article-title":"Interplay of ferroptotic and apoptotic cell death and its modulation by BH3-mimetics","volume":"32","author":"Qiu","year":"2025","journal-title":"Cell Death Differ."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"743006","DOI":"10.3389\/fonc.2021.743006","article-title":"Molecular Mechanisms of Ferroptosis and Its Roles in Hematologic Malignancies","volume":"11","author":"Zhao","year":"2021","journal-title":"Front. Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1177\/1535370215593826","article-title":"Galectin-3 in autoimmunity and autoimmune diseases","volume":"240","author":"Gatto","year":"2015","journal-title":"Exp. Biol. Med."},{"key":"ref_33","first-page":"9247574","article-title":"The Many Roles of Galectin-3, a Multifaceted Molecule, in Innate Immune Responses against Pathogens","volume":"2017","author":"Ortega","year":"2017","journal-title":"Mediat. Inflamm."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1626","DOI":"10.1016\/j.celrep.2015.02.012","article-title":"Microglia-Secreted Galectin-3 Acts as a Toll-like Receptor 4 Ligand and Contributes to Microglial Activation","volume":"10","author":"Burguillos","year":"2015","journal-title":"Cell Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e18798","DOI":"10.1097\/MD.0000000000018798","article-title":"Renal hemosiderosis secondary to intravascular hemolysis after mitral valve repair: A case report","volume":"99","author":"Lee","year":"2020","journal-title":"Medicine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"107496","DOI":"10.1016\/j.isci.2023.107496","article-title":"Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization","volume":"26","author":"Liu","year":"2023","journal-title":"iScience"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1111\/j.1600-065X.2009.00794.x","article-title":"The regulation of inflammation by galectin-3","volume":"230","author":"Henderson","year":"2009","journal-title":"Immunol. Rev."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1093\/toxsci\/kfac057","article-title":"Recovered Hepatocytes Promote Macrophage Apoptosis Through CXCR4 After Acetaminophen-Induced Liver Injury in Mice","volume":"188","author":"Nguyen","year":"2022","journal-title":"Toxicol. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1016\/j.cell.2010.01.040","article-title":"The inflammasomes","volume":"140","author":"Schroder","year":"2010","journal-title":"Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1964","DOI":"10.3389\/fimmu.2020.01964","article-title":"TLR4 Signaling by Heme and the Role of Heme-Binding Blood Proteins","volume":"11","author":"Janciauskiene","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1016\/j.kint.2024.06.023","article-title":"Galectin-3 contributes to pathogenesis of IgA nephropathy","volume":"106","author":"Chou","year":"2024","journal-title":"Kidney Int."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e2026246118","DOI":"10.1073\/pnas.2026246118","article-title":"Galectin-3 promotes noncanonical inflammasome activation through intracellular binding to lipopolysaccharide glycans","volume":"118","author":"Lo","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1093\/ckj\/sfx121","article-title":"Clinical profile and outcome of pigment-induced nephropathy","volume":"11","author":"Sakthirajan","year":"2018","journal-title":"Clin. Kidney J."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.toxlet.2005.03.004","article-title":"Free heme toxicity and its detoxification systems in human","volume":"157","author":"Kumar","year":"2005","journal-title":"Toxicol. Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1038\/labinvest.3780131","article-title":"Galectin-1 and galectin-3 in chronic pancreatitis","volume":"80","author":"Wang","year":"2000","journal-title":"Lab. Investig."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"6978936","DOI":"10.1155\/2016\/6978936","article-title":"The Role of Toll-Like Receptor 4 in Infectious and Noninfectious Inflammation","volume":"2016","author":"Molteni","year":"2016","journal-title":"Mediat. Inflamm."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Cenariu, D., Iluta, S., Zimta, A.A., Petrushev, B., Qian, L., Dirzu, N., Tomuleasa, C., Bumbea, H., and Zaharie, F. (2021). Extramedullary Hematopoiesis of the Liver and Spleen. J. Clin. Med., 10.","DOI":"10.3390\/jcm10245831"},{"key":"ref_48","first-page":"211","article-title":"Heme sensitization to TNF-mediated programmed cell death","volume":"Volume 691","author":"Gozzelino","year":"2011","journal-title":"Advances in TNF Family Research, Proceedings of the 12th International TNF Conference, Madrid, Spain, 26\u201329 April 2009"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"14496","DOI":"10.1073\/pnas.0903497106","article-title":"Galectin-3 negatively regulates TCR-mediated CD4+ T-cell activation at the immunological synapse","volume":"106","author":"Chen","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7228","DOI":"10.1158\/0008-5472.CAN-08-1245","article-title":"Tumor-associated galectin-3 modulates the function of tumor-reactive T cells","volume":"68","author":"Peng","year":"2008","journal-title":"Cancer Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2650","DOI":"10.4049\/jimmunol.180.4.2650","article-title":"Regulation of alternative macrophage activation by galectin-3","volume":"180","author":"MacKinnon","year":"2008","journal-title":"J. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1517\/14728222.6.4.461","article-title":"Galectins: Novel anti-inflammatory drug targets","volume":"6","author":"Liu","year":"2002","journal-title":"Expert Opin. Ther. Targets"}],"container-title":["Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-9721\/14\/5\/161\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T10:02:49Z","timestamp":1778061769000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-9721\/14\/5\/161"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,6]]},"references-count":52,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["diseases14050161"],"URL":"https:\/\/doi.org\/10.3390\/diseases14050161","relation":{},"ISSN":["2079-9721"],"issn-type":[{"value":"2079-9721","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,6]]}}}