{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T01:31:54Z","timestamp":1789781514352,"version":"4.0.1"},"reference-count":42,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2017,1,10]],"date-time":"2017-01-10T00:00:00Z","timestamp":1484006400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Local accumulation of Advanced Oxidation Protein Products (AOPP) induces pro-inflammatory and pro-fibrotic processes in kidneys and is an independent predictor of renal fibrosis and of rapid decline of eGFR in patients with chronic kidney disease (CKD). In addition to kidney damage, circulating AOPP may be regarded as mediators of systemic oxidative stress and, in this capacity, they might play a role in the progression of atherosclerotic damage of arterial walls. Atherosclerosis is a chronic inflammatory disease that involves activation of innate and adaptive immunity. Dendritic cells (DCs) are key cells in this process, due to their role in antigen presentation, inflammation resolution and T cell activation. AOPP consist in oxidative modifications of proteins (such as albumin and fibrinogen) that mainly occur through myeloperoxidase (MPO)-derived hypochlorite (HOCl). HOCl modified proteins have been found in atherosclerotic lesions. The oxidizing environment and the shifts in cellular redox equilibrium trigger inflammation, activate immune cells and induce immune responses. Thus, surface thiol groups contribute to the regulation of immune functions. The aims of this work are: (1) to evaluate whether AOPP-proteins induce activation and differentiation of mature macrophages into dendritic cells in vitro; and (2) to define the role of cell surface thiol groups and of free radicals in this process. AOPP-proteins were prepared by in vitro incubation of human serum albumin (HSA) with HOCl. Mouse macrophage-like RAW264.7 were treated with various concentrations of AOPP-HSA with or without the antioxidant N-acetyl cysteine (NAC). Following 48 h of HSA-AOPP treatment, RAW264.7 morphological changes were evaluated by microscopic observation, while markers of dendritic lineage and activation (CD40, CD86, and MHC class II) and allogeneic T cell proliferation were evaluated by flow cytometry. Cell surface thiols were measured by AlexaFluor-maleimide binding, and ROS production was assessed as DCF fluorescence by flow cytometry. HSA-AOPP induced the differentiation of RAW264.7 cells into a dendritic-like phenotype, as shown by morphological changes, by increased CD40, CD86 and MHC class II surface expression and by induction of T cell proliferation. The cell surface thiols dose dependently decreased following HSA-AOPP treatment, while ROS production increased. NAC pre-treatment enhanced the amount of cell surface thiols and prevented their reduction due to treatment with AOPP. Both ROS production and RAW264.7 differentiation into DC-like cells induced by HSA-AOPP were reduced by NAC. Our results highlight that oxidized plasma proteins modulate specific immune responses of macrophages through a process involving changes in the thiol redox equilibrium. We suggest that this mechanism may play a role in determining the rapid progression of the atherosclerotic process observed in CKD patients.<\/jats:p>","DOI":"10.3390\/toxins9010027","type":"journal-article","created":{"date-parts":[[2017,1,10]],"date-time":"2017-01-10T10:16:18Z","timestamp":1484043378000},"page":"27","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Advanced Oxidation Protein Products-Modified Albumin Induces Differentiation of RAW264.7 Macrophages into Dendritic-Like Cells Which Is Modulated by Cell Surface Thiols"],"prefix":"10.3390","volume":"9","author":[{"given":"Silvano","family":"Garibaldi","sequence":"first","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Barisione","sequence":"additional","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3056-6569","authenticated-orcid":false,"given":"Barbara","family":"Marengo","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pietro","family":"Ameri","sequence":"additional","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudio","family":"Brunelli","sequence":"additional","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manrico","family":"Balbi","sequence":"additional","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giorgio","family":"Ghigliotti","sequence":"additional","affiliation":[{"name":"Division of Cardiology, IRCCS University Hospital San Martino, Research Centre of Cardiovascular Biology, University of Genova, Genova 16132, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1038\/ki.1996.186","article-title":"Advanced oxidation protein products as a novel marker of oxidative stress in uremia","volume":"49","author":"Friedlander","year":"1996","journal-title":"Kidney Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.bbadis.2004.02.008","article-title":"Biochemical and spectrophotometric significance of advanced oxidized protein products","volume":"1689","author":"Gausson","year":"2004","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1310","DOI":"10.1093\/ndt\/12.7.1310","article-title":"Advanced oxidation protein products: Novel uraemic toxins and pro-inflammatory mediators in chronic renal failure?","volume":"12","year":"1997","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_4","first-page":"S11","article-title":"Are advanced oxidation protein products potential uremic toxins?","volume":"84","author":"Gausson","year":"2003","journal-title":"Kidney Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/S0021-9150(01)00706-7","article-title":"Increased level of advanced oxidation protein products in patients with coronary artery disease","volume":"162","author":"Kaneda","year":"2002","journal-title":"Atherosclerosis"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/S1262-3636(07)70230-6","article-title":"Protein oxidation parameters in type 2 diabetic patients with good and poor glycaemic control","volume":"31","author":"Cakatay","year":"2005","journal-title":"Diabetes Metab."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1829","DOI":"10.1210\/en.2007-1544","article-title":"Advanced oxidation protein products promote inflammation in diabetic kidney through activation of renal nicotinamide adenine dinucleotide phosphate oxidase","volume":"149","author":"Shi","year":"2008","journal-title":"Endocrinology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1136\/ard.2006.067504","article-title":"Radical oxygen species production induced by advanced oxidation protein products predicts clinical evolution and response to treatment in systemic sclerosis","volume":"66","author":"Servettaz","year":"2007","journal-title":"Ann. Rheum. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1053\/j.ajkd.2004.09.011","article-title":"Advanced oxidation protein products as risk factors for atherosclerotic cardiovascular events in nondiabetic predialysis patients","volume":"45","author":"Nguyen","year":"2005","journal-title":"Am. J. Kidney Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"665","DOI":"10.3390\/toxins6020665","article-title":"Protein-bound uremic toxins: New culprits of cardiovascular events in chronic kidney disease patients","volume":"6","author":"Ito","year":"2014","journal-title":"Toxins"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1046\/j.1523-1755.2003.00044.x","article-title":"AOPP-induced activation of human neutrophil and monocyte oxidative metabolism: A potential target for N-acetylcysteine treatment in dialysis patients","volume":"64","author":"Gausson","year":"2003","journal-title":"Kidney Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2524","DOI":"10.4049\/jimmunol.161.5.2524","article-title":"Advanced oxidation protein products as novel mediators of inflammation and monocyte activation in chronic renal failure","volume":"161","author":"Friedlander","year":"1998","journal-title":"J. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1146\/annurev-immunol-020711-074950","article-title":"The dendritic cell lineage: Ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting","volume":"31","author":"Merad","year":"2013","journal-title":"Annu. Rev. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2535","DOI":"10.1002\/eji.201242480","article-title":"Role of inflammatory dendritic cells in innate and adaptive immunity","volume":"42","author":"Hespel","year":"2012","journal-title":"Eur. J. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"257","DOI":"10.17219\/acem\/40463","article-title":"Differentiation of Monocyte Derived Dendritic Cells in End Stage Renal Disease is Skewed towards Accelerated Maturation","volume":"24","author":"Dopheide","year":"2015","journal-title":"Adv. Clin. Exp. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1286","DOI":"10.1172\/JCI38399","article-title":"Kidney dendritic cell activation is required for progression of renal disease in a mouse model of glomerular injury","volume":"119","author":"Heymann","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2185","DOI":"10.1093\/ndt\/gfu076","article-title":"Dendritic cells in progressive renal disease: Some answers, many questions","volume":"29","author":"Kitching","year":"2014","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.freeradbiomed.2013.05.050","article-title":"The thiol pool in human plasma: The central contribution of albumin to redox processes","volume":"65","author":"Turell","year":"2013","journal-title":"Free Radic. Biol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/S0092-8674(01)00449-4","article-title":"Dendritic cells: Specialized and regulated antigen processing machines","volume":"106","author":"Mellman","year":"2001","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/j.cmet.2010.09.010","article-title":"Atherogenic lipids and lipoproteins trigger CD36-TLR2-dependent apoptosis in macrophages undergoing endoplasmic reticulum stress","volume":"12","author":"Seimon","year":"2010","journal-title":"Cell Metab."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1046\/j.1523-1755.2001.00807.x","article-title":"Albumin is the major plasma protein target of oxidant stress in uremia","volume":"60","author":"Himmelfarb","year":"2001","journal-title":"Kidney Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5465","DOI":"10.1016\/j.bbagen.2013.04.036","article-title":"Redox properties of serum albumin","volume":"1830","author":"Anraku","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1021\/tx800372d","article-title":"What Are the Plasma Targets of the Oxidant Hypochlorous Acid? A Kinetic Modeling Approach","volume":"22","author":"Pattison","year":"2009","journal-title":"Chem. Res. Toxicol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/j.clinbiochem.2011.03.134","article-title":"Role of advanced oxidation protein products and Thiol ratio in patients with acute coronary syndromes","volume":"44","author":"Barsotti","year":"2011","journal-title":"Clin. Biochem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1016\/j.atherosclerosis.2007.11.030","article-title":"Postprandial serum induces apoptosis in endothelial cells: Role of polymorphonuclear-derived myeloperoxidase and metalloproteinase-9 activity","volume":"198","author":"Spallarossa","year":"2008","journal-title":"Atherosclerosis"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Biedro\u0144, R., Konopi\u0144ski, M.K., Marcinkiewicz, J., and J\u00f3zefowski, S. (2015). Oxidation by neutrophils-derived HOCl increases immunogenicity of proteins by converting them into ligands of several endocytic receptors involved in antigen uptake by dendritic cells and macrophages. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0123293"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1681\/ASN.2011070680","article-title":"The renal mononuclear phagocytic system","volume":"23","author":"Nelson","year":"2012","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1038\/ki.2011.129","article-title":"Homeostatic and pathogenic role of renal dendritic cells","volume":"80","author":"Teteris","year":"2011","journal-title":"Kidney Int."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/S0967-2109(00)00141-1","article-title":"CD40 co-stimulatory molecule expression by dendritic cells in primary atherosclerotic lesions in carotid arteries and in stenotic saphenous vein coronary artery grafts","volume":"9","author":"Ozmen","year":"2001","journal-title":"Cardiovasc. Surg."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/j.ijcard.2004.05.022","article-title":"Differentiation of dendritic cells in monocyte cultures isolated from patients with unstable angina","volume":"97","author":"Sharma","year":"2004","journal-title":"Int. J. Cardiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/S0891-5849(01)00735-3","article-title":"The role of advanced oxidation protein products in regulation of dendritic cell function","volume":"32","author":"Alderman","year":"2002","journal-title":"Free Radic. Biol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4001","DOI":"10.1073\/pnas.2628032100","article-title":"Lymphocyte surface thiol levels","volume":"100","author":"Sahaf","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"871","DOI":"10.2131\/jts.35.871","article-title":"Changes of cell-surface thiols and intracellular signaling in human monocytic cell line THP-1 treated with diphenylcyclopropenone","volume":"35","author":"Hirota","year":"2010","journal-title":"J. Toxicol. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1096\/fj.02-0105fje","article-title":"Glutathione disulfide induces apoptosis in U937 cells by a redox-mediated p38 MAP kinase pathway","volume":"17","author":"Filomeni","year":"2003","journal-title":"FASEB J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/j.freeradbiomed.2005.01.002","article-title":"Redox regulation of the signaling pathways leading to eNOS phosphorylation","volume":"38","author":"Tanaka","year":"2005","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1016\/j.jacc.2005.09.063","article-title":"The relationship between plasma levels of oxidized and reduced thiols and early atherosclerosis in healthy adults","volume":"47","author":"Ashfaq","year":"2006","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2973","DOI":"10.1161\/CIRCULATIONAHA.104.515155","article-title":"Intracellular proatherogenic events and cell adhesion modulated by extracellular thiol\/disulfide redox state","volume":"111","author":"Go","year":"2005","journal-title":"Circulation"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"724","DOI":"10.1055\/s-0037-1615659","article-title":"N-acetyl-cysteine reduces neointimal thickening and procoagulant activity after balloon-induced injury in abdominal aortae of New Zealand white rabbits","volume":"85","author":"Ghigliotti","year":"2001","journal-title":"Thromb. Haemost."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1016\/S0735-1097(97)00005-3","article-title":"Effect of transdermal nitroglycerin or N-acetylcysteine, or both, in the long-term treatment of unstable angina pectoris","volume":"29","author":"Demicheli","year":"1997","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1089\/ars.2012.4603","article-title":"Advanced oxidation protein products activate intrarenal renin-angiotensin system via a CD36-mediated, redox-dependent pathway","volume":"18","author":"Cao","year":"2013","journal-title":"Antioxid. Redox Signal"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1016\/j.cca.2012.01.038","article-title":"Improved detection of advanced oxidation protein products in plasma","volume":"413","author":"Hanasand","year":"2012","journal-title":"Clin. Chim. Acta"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Colla, R., Izzotti, A., De Ciucis, C., Fenoglio, D., Ravera, S., Speciale, A., Ricciarelli, R., Furfaro, A.L., Pulliero, A., and Passalacqua, M. (2016). Glutathione-mediated antioxidant response and aerobic metabolism: Two crucial factors involved in determining the multi-drug resistance of high-risk neuroblastoma. Oncotarget.","DOI":"10.18632\/oncotarget.12209"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/9\/1\/27\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:25:51Z","timestamp":1760207151000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/9\/1\/27"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,1,10]]},"references-count":42,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,1]]}},"alternative-id":["toxins9010027"],"URL":"https:\/\/doi.org\/10.3390\/toxins9010027","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,1,10]]}}}