Summary
A highly sensitive method of ultrastructural-immunoperoxidase staining was developed for use with monoclonal antibodies which have been raised in this laboratory to a variety of antigens of the human kidney. Because of the human kidney. Because of the susceptibility of the antigens to fixation and processing, a four layer, pre-embedding method of staining was used. Results confirmed and clarified previously reported light microscopy results, indicating that an antigen recognized by the PHM5 antibody was found on the podocyte cell membrane within the glomerulus and was not present within the glomerular basement membrane. The antigen was also present on the extraglomerular endothelial cell membrane. The study also demonstrated the presence of an antigen specific to endothelial cells throughout the renal cortex, and gave further insight into the precise localization of glomerular basement membrane components including fibronectin. The method of staining is now being used together with detailed ultrastructural studies to identify the cells produced from isolated glomeruli in tissue culture.
Similar content being viewed by others
References
Avrameas, S. &Ternynck, T. (1971) Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration.Immunochemistry 8, 1175–9.
Becker, G., Hancock, W. W., Karaft, N. &Atkins, R. C. (1981) Monoclonal antibodies to human macrophage and leucocyte common antigens.Pathology 13, 669–80.
Courtoy, P. J., Kanwar, Y. S., Hynes, R. O. &Farquhar, M. G. (1980) Fibronectin localisation in the rat glomerulus.J. Cell Biol. 87, 691–6.
Courtoy, P. J., Picton, D. H. &Farquhar, M. G. (1983) Resolution and limitations of the immunoperoxidase procedure in the localisation of extracellular matrix antigens.J. Histochem. Cytochem. 31, 945–51.
Courtoy, P. J., Timpl, R. &Farquhar, M. G. (1982) Comparative distribution of laminin Type IV collagen and fibronectin in the rat glomerulus.J. Histochem. Cytochem. 30, 874–86.
Glasgow, E. E., Hancock, W. &Atkins, R. C. (1981) The technique of glomerular culture. InTechniques for Nephropathology (edited byAllen, D. &Dowling, J. P.) Boca Raton: CRC Press pp. 87–93.
Graham, R. C. Jr. &Karnovsky, M. J. (1964) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of the mouse kidney. Ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14, 291–302.
Hancock, W. &Atkins, R. C. (1981) Monoclonal antibodies to human renal antigens.Aust. N. Z. Med. 11, 594.
Hancock, W. &Atkins, R. C. (1983) Monoclonal antibodies to human glomerular cells: A marker for glomerular epithelial cells.Nephron 33, 83–90.
Hancock, W., Becker, G. J. &Atkins, R. C. (1982) A comparison of fixatives and immunohistological technics for use with monoclonal antibodies to cell surface antigens.Am. J. Clin. Path. 78(6), 825–31.
Hancock, W., Kraft, N., Clarke, F. &Atkins, R. C. (1984) Production of monoclonal antibodies to fibronectin, Type IV collagen and other antigens of the human glomerulus.Pathology 16, 197–206.
Jackson, A. E., Glasgow, E. F. &Atkins, R. C. (1989) Ultrastructural immunoperoxidase investigations of human glomerular cells in tissue culture.Nephron 52, 62–71.
Kraehenbuhl, J. P. &Jamieson, J. D. (1971) Localisation in intracellular antigens by immunoelectron microscopy.Int. Rev. Exper. Pathol. 13, 1–53.
McLean, I. W. &Nakane, P. K. (1974) Periodate-Lysineparaformaldehyde fixative. A new fixative for immunoelectron microscopy.J. Histochem. Cytochem. 22, 1077–83.
Martinez-Hernandez, A., Marsh, C. A., Clark, C. C., Macarak, E. J. &Brownell, A. G. (1981) Fibronectin: Its relationship to basement membranes. II. Ultrastructural studies in rat kidney.Coll. Res. 5, 405–18.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jackson, A.E., Atkins, R.C. & Glasgow, E.F. Ultrastructural localization of human kidney antigens using monoclonal antibodies. Histochem J 23, 509–516 (1991). https://doi.org/10.1007/BF01041177
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01041177