Immunohistochemical localization of different epitopes of advanced glycation end products in human atherosclerotic lesions

Atherosclerosis. 1998 Nov;141(1):61-75. doi: 10.1016/s0021-9150(98)00149-x.


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.

MeSH terms

  • Adult
  • Aged
  • Antibodies, Monoclonal
  • Aorta / chemistry*
  • Aorta / pathology
  • Arteriosclerosis / metabolism*
  • Arteriosclerosis / pathology
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / analysis*
  • Extracellular Matrix / chemistry
  • Female
  • Glycation End Products, Advanced / analysis*
  • Glycation End Products, Advanced / immunology
  • Humans
  • Immunohistochemistry
  • Lipoproteins, LDL / metabolism
  • Lysine / analogs & derivatives
  • Lysine / analysis
  • Lysine / immunology
  • Macrophages / chemistry
  • Male
  • Middle Aged
  • Tunica Intima / chemistry


  • Antibodies, Monoclonal
  • Epitopes
  • Glycation End Products, Advanced
  • Lipoproteins, LDL
  • oxidized low density lipoprotein
  • N(6)-carboxymethyllysine
  • Lysine