title: Immunohistochemical localization of different epitopes of advanced glycation end products in human atherosclerotic lesions.
abstract: To better understand the role of advanced glycation end products (AGEs) in atherogenesis, we developed specific antibodies against different immunological epitopes of AGE structures, including Nepsilon-(carboxymethyl)lysine-protein adduct (CML) and a structure(s) other than CML (nonCML), and demonstrated the immunohistochemical localization of CML- and nonCML-epitopes in atherosclerotic lesions of human aorta, which were obtained at autopsy from 20 nondiabetic patients (12 males and eight females; mean age, 60.8+/-16.7 years). Monoclonal anti-CML antibody (6D12) recognized not only AGE-modified proteins, but also CML-modified proteins. On the other hand, polyclonal anti-nonCML antibody reacted to AGE-modified proteins, but not to CML-modified proteins. Both antibodies were unreactive to the early-stage products of glycation, including fructose-modified butyloxycarbonyl-lysine and fructose-epsilon-aminocaproic acid. Atherosclerotic lesions included diffuse intimal thickening (DIT), fatty streaks (FS), atherosclerotic plaques (AP) and complicated lesions. An immunohistochemical analysis showed both CML- and nonCML-epitopes to be found along the collagen fibers in DIT in subjects more than 40 years old, but not in subjects less than 40 years old. CML-epitopes accumulated mainly in the cytoplasm of macrophage/foam cells, while nonCML-epitopes accumulated exclusively in the extracellular spaces in FS. APs showed the CML-epitope stored macrophage/foam cells, and the accumulation of both CML- and nonCML-epitopes in the lipid-rich fibrous area. An immunohistochemical analysis with a monoclonal antibody against oxidized low density lipoprotein (FOH1a/DLH3) showed the presence of this antigen within the cytoplasm of the macrophage/foam cells in atherosclerotic lesions, which were also positive for the CML-epitopes. These findings thus suggest that the heterogeneous localization of AGEs in atherosclerotic lesions depends on their different epitopes, and that a close link, therefore, exists between the peroxidation of LDL and the formation of AGEs in atherosclerotic lesions.
authors.affiliation: Second Department of Pathology, School of Medicine, Fukuoka University, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Sakata
authors.forename: Y
authors.initials: Y
authors.lastname: Imanaga
authors.forename: J
authors.initials: J
authors.lastname: Meng
authors.forename: Y
authors.initials: Y
authors.lastname: Tachikawa
authors.forename: S
authors.initials: S
authors.lastname: Takebayashi
authors.forename: R
authors.initials: R
authors.lastname: Nagai
authors.forename: S
authors.initials: S
authors.lastname: Horiuchi
authors.forename: H
authors.initials: H
authors.lastname: Itabe
authors.forename: T
authors.initials: T
authors.lastname: Takano
mesh_terms: D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D001011:Aorta / Q000737:chemistry* / Q000473:pathology; D001161:Arteriosclerosis / Q000378:metabolism* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005109:Extracellular Matrix / Q000737:chemistry; D005260:Female; D017127:Glycation End Products, Advanced / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives / Q000032:analysis / Q000276:immunology; D008264:Macrophages / Q000737:chemistry; D008297:Male; D008875:Middle Aged; D017539:Tunica Intima / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D017127:Glycation End Products, Advanced; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; C048496:N(6)-carboxymethyllysine; D008239:Lysine