Immunohistochemical detection of hypochlorite-modified proteins in glomeruli of human membranous glomerulonephritis.
A proposed analogy between atherosclerosis and glomerulosclerosis suggests that factors that contribute to the development of atherosclerosis, ie, oxidatively modified (lipo)proteins, may also participate in glomerular injury. Although the nature of the in vivo oxidants has not been clearly identified, increasing evidence suggested the myeloperoxidase (MPO)-H(2)O(2)-halide system to be responsible for the damage observed in leukocyte-dependent glomerulonephritis. MPO, a heme protein secreted by activated phagocytes, may generate modified/oxidized proteins in vivo via intermediate formation of hypochlorous acid (HOCl)/hypochlorite. HOCl, a reactive oxygen species and powerful oxidant, can convert (lipo)proteins into atherogenic forms in vitro and in vivo. Here we demonstrate the presence of HOCl-modified proteins in glomeruli of patients with membranous glomerulonephritis using monoclonal antibodies that do not cross-react with other oxidative modifications. Immunostaining for HOCl-modified epitopes in human minimal change glomerulopathy revealed glomeruli that were unreactive, although the number of MPO-positive cells/glomerulus was slightly increased in comparison to controls. In contrast to minimal change glomerulopathy, a pronounced infiltration of mononuclear cells/glomerulus in membranoproliferative glomerulonephritis is in line with pronounced staining for HOCl-modified epitopes. Immunostaining was detected in intracapillary cells and immune complex deposits within the glomerular basement membrane. In human membranous glomerulonephritis (Stages I to III), staining for HOCl-modified proteins was localized at the basement membrane and podocytes. Staining of serial sections revealed colocalization of HOCl-modified epitopes and MPO in glomerular peripheral basement membranes. Subsequently, tubulointerstitial staining for HOCl-modified epitopes was observed in foam cells at the border of the cytoplasm and in damaged tubular epithelia in focal advanced chronic lesions. Our results indicate that oxidative modification of the basement membrane structure by phagocyte-derived HOCl may be of importance for glomerular defects. The observed colocalization of HOCl-modified proteins and MPO in podocytes and adjacent basement membranes strengthens the assumption that the MPO-H(2)O(2)-halide system contributes to glomerular dysfunction in patients with membranous glomerulonephritis.
Gröne
Hermann-Josef
HJ
Department of Cellular and Molecular Pathology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany. h-j.groene@dkfz-heidelberg.de
Gröne
Elisabeth F
EF
Malle
Ernst
E
D000911:Antibodies, Monoclonal; D001707:Biopsy, Needle; D015432:Glomerulonephritis, Membranoproliferative / Q000473:pathology; D015433:Glomerulonephritis, Membranous / Q000473:pathology*; D006801:Humans; D006997:Hypochlorous Acid; D007150:Immunohistochemistry / Q000379:methods; D007202:Indicators and Reagents; D007668:Kidney / Q000166:cytology / Q000473:pathology*; D009195:Peroxidase / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012016:Reference Values
D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D011506:Proteins; D006997:Hypochlorous Acid; D009195:Peroxidase