Summary
The ultrastructural localization of immunoglobulins (Igs) in immunologically mediated glomerulonephritis was achieved in ultrathin sections of glutaraldehyde-fixed, post-osmicated, Epon-embedded human renal needle biopsies using the indirect immunogold staining procedure. Extracellular Igs were identified in electron-dense deposits in subendothelia, intramembranous, subepithelial and mesangial locations in different cases of immune-complex-mediated glomerulonephritis. The excellent preservation of the ultrastructural morphology of the tissue sections made it possible to precisely correlate the presence and distribution of the antigens with those of the electron-dense deposits, which are the ultrastructural hallmarks of these diseases. Our post-embedding staining procedure which allows extracellular Igs to be accurately localized should be a very useful tool for immunomorphological investigations of glomerulo-nephritis.
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References
Coggi G, Dell'Orto P, Grigolato PG, Sacchi G, Viale G (1985) Immunoelectron microscopy of human renal biopsies: prerequisites and limitations. Appl Pathol (in press)
Davies DR, Tighe JR, Wing AJ, Jones NF (1977) Immunoglobulin deposition in membranous glomerulonephritis: immunofluorescence and immunoelectron microscopy findings. Histopathology 1:39–52
Dell'Orto P, Viale G, Colombi R, Braidotti P, Coggi G (1982) Immunohistochemical localization of human immunoglobulins and lysozyme in epoxy-embedded lymph nodes. Effect of different fixatives and of proteolytic digestion. J Histochem Cytochem 30:630–636
De Mey J, Moeremans M, Geuens G, Nuydens R, De Brabander R (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immunogold staining) method. Cell Biol Int Rep 5:889–899
Doi T, Kanatsu K, Nagai H, Kohrogi N, Kuwahara T, Hamashima Y (1984) Immunoelectron microscopic studies of IgA nephropathy. Nephron 36:246–251
Dysart NK Jr, Sisson S, Vernier RL (1983) Immunoelectron microscopy of IgA nephropathy. Clin Immunol Immunopathol 29:254–270
Horisberger M, Rosset J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25:295–305
Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. A comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Laliberté F, Druet P, Bellon B, Sapin C, Bariéty J (1974) Application des tecyniques d'immunopéroxydase a l'étude ultrastructurale des glomérulonéphrites. J Microsc 21:283–288
Leduc EH, Scott GB, Avrameas S (1969) Ultrastructural localization of intracellular immune globulins in plasma cells and lymphoblats by enzyme-labeled antibodies. J Histochem Cytochem 17:211–224
Rantala I, Helin H (1982) Immunoelectron microscopic localization of immunoglobulins and complement in human renal glomeruli. J Histochem Cytochem 30:932–937
Rodning CB, Erlandsen SL, Coulter HD, Wilson ID (1980a) Immunohistochemical localization of IgA antigens in sections embedded in Epoxy resin. J Histochem Cytochem 28:199–205
Rodning CB, Erlandsen SL, Wilson ID (1980b) Immunohistochemical identification of immunoglobulin A on ultrathin tissue sections. Am J Anat 157:221–224
Rossmann P, Jirka J, Matousovic K, Zastava V, Sefcova J (1984) Ultrastructural immunohistochemistry of glomerulonephritis in needle biopsy. Pathol Res Pract 178:251–260
Shindo N, Kobayashi E, Okada M (1984) Immunoelectron microscopic (IEM) studies on glutaraldehyde-fixed renal specimen. J Histochem Cytochem 32:501–509
Slusarczyk J, Michalak T, Nazarewicz de Mezer T, Krawczynski K, Nowoslawski A (1980) Membranous glomerulopathy associated with hepatitis B core antigen immune complexes in children. Am J Pathol 98:29–44
Smit JW, Meijer CJLM, Decary F, Feltkamp-Vroom TM (1974) Paraformaldehyde fixation in immunofluorescence and immunoelectron microscopy. Preservation of tissue and cell surface membrane antigen. J Immunol Methods 6:93–98
Takamiya H, Batsford SR, Vogt A (1980) An approach to postem-bedding staining of protein (immunoglobulin) antigen embedded in plastic: prerequisites and limitations. J Histochem Cytochem 28:1041–1049
Tapia FJ, Varndell IM, Probert L, De Mey J, Polak JM (1983) Double immunogold staining method for the simultaneous ultrastructural localization of regulatory peptides. J Histochem Cytochem 31:977–981
Viale G, Dell'Orto P, Braidotti P, Coggi G (1985) Ultrastructural localization of intracellular immunoglobulins in Epon-embedded human lymph nodes. An immunoelectron microscopic investigation using the immunogold staining (IGS) and the avidin-biotin-peroxidase complex (ABC) methods. J Histochem Cytochem 33:400–406
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Viale, G., Dell'Orto, P., Colombi, R. et al. Ultrastructural localization of extracellular immunoglobulins in immune-complex-mediated glomerulonephritis. Histochemistry 84, 1–4 (1986). https://doi.org/10.1007/BF00493411
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DOI: https://doi.org/10.1007/BF00493411