Summary
Antibodies to α-elastin peptides, amyloid P component, lysozyme and plasma protease, inhibitors have been used in an immunoperoxidase method to stain elastic fibres in frozen sections of human breast tissues. A loss of immunoreactivity seen in formalin-fixed, paraffin-embedded sections was reversed by a preliminary proteolysis. Differences in the tinctorial dye and immunohistochemical staining patterns following proteolysis by a variety of enzymes suggests a selective unmasking or removal of elastic fibre components and thus the presence of separate binding sites for individual antibodies and tinctorial dyes. Antibody blocking experiments and double immunoenzymatic labelling support the existence of several different epitopes within elastic fibres.
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Barnard, K., Davies, J. D. &Young, E. W. (1982a) Immunoperoxidase localization of elastin in the human aorta.Experientia 38, 984–5.
Barnard, K., Partridge, S. M., Whiting, A. H., Fantl, V. &McCullagh, K. G. (1982b) Immunologic studies with human aortic elastin.Conn. Tiss. Res. 9, 233–40.
Bartholomew, J. S. &Anderson, J. C. (1983) Investigation of relationships between collagens, elastin and proteoglycans in bovine thoracic aorta by immunofluorescence techniques.Histochem. J. 15, 1177–90.
Bodley, H. D. &Wood, R. L. (1972) Ultrastructural studies on elastic fibres using enzymatic digestion of thin sections.Anat. Rec. 172, 71–88.
Breathnach, S. M., Melrose, S. M., Bhogal, B., De Beer, F. C., Black, M. M. &Pepys, M. B. (1982) Immunohistochemical studies of amyloid P component in disorders of cutaneous elastic tissue.Br. J. Dermat. 107, 443–52.
Breathnach, S. M., Melrose, S. M., Bhogal, B., De Beer, F. C., Dyck, R. F., Tennent, G., Black, M. M. &Pepys, M. B. (1981) Amyloid P component is located on elastic fibre microfibrils in normal human tissues.Nature 293, 652–3.
Curran, R. C. &Gregory, J. (1978) Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques.J. clin. Path. 31, 974–83.
Davies, J. D. (1973) Hyperelastosis, obliteration and fibrous plaques in major ducts of the human breast.J. Path. 110, 13–26.
Davies, J. D., Barnard, K., Whicher, J. T. &Young E. W. (1983a) Breast elastosis: association with lysozyme and certain protease inhibitors.J. Path. 139, 477.
Davies, J. D. Barnard, K. &Young, E. W. (1982) Immunoreactive elastin in benign breast tissues.Virchows Arch. path. Anat. Physiol. 398, 109–17.
Davies, J. D. &Young, E. W. (1982) Congo blue: A rapid stain for elastic fibres.J. clin. Path. 35, 789–91.
Davies, J. D., Young, E. W., Mera, S. L. &Barnard, K. (1983b) Lysozyme is a component of human vascular elastic fibres.Experientia 39, 382–3.
Fischer, E. R., Rosenthal, T. B. &Lansing, A. I. (1960) Elastolytic effect of pepsin.J. Histochem. Cytochem. 8, 102–4.
Fleischmajer, R. &Timpl, R. (1984) Ultrastructural localization of fibronectin to different anatomic structures of human skin.J. Histochem. Cytochem. 32, 315–21.
Horobin, R. W. (1982)Histochemistry. London: Butterworths.
Jones, C. J., Sear, C. H. &Grant, M. E. (1980) An ultrastructural study of fibroblasts derived from bovine ligamentum nuchae and their capacity for elastogenesis in culture.J. Path. 131, 35–53.
Kadar, A. (1977) Scanning electron microscopy of purified elastin with and without enzymatic digestion.Adv. exp. Med. Biol. 79, 71–96.
Kadar, A. (1979) The elastic fiber: normal and pathological conditions in the arteries.Expl Path. (Jena) Suppl. 5, 9–130.
Kadar, A. (1980) New concepts of elastic tissue disorders. InFrontiers of Matrix, Biology, Vol. 8, (edited byRobert, L.), pp. 54–68. Basel: Krager.
Kadar, A., Gardner, D. L. &Bush, V. (1971) Susceptibility of the chick-embryo aorta to elastase: An electron microscope study.J. Path. 104, 261–6.
McCullagh, K. G., Barnard, K., Davies, J. D. &Partridge, S. M. (1980) Newly synthesised elastin is on the surface of tumour cells in human mammary carcinoma.Experientia 36, 1315–16.
Mangkornkanok-Mark, M., Eisenstein, R. &Bahu, R. M. (1981) Immunological studies of bovine aortic and cartilage proteoglycans.J. Histochem. Cytochem. 29, 547–52.
Mason, D. Y., Abdulaziz, Z., Falini, B. &Stein, H. (1983) Double immunoenzymatic labelling. InImmunocytochemistry (edited byPolak, J. M. andVan Noorden, S.), pp. 113–28. Bristol: John Wright and Sons.
Mera, S. L. &Davies, J. D. (1984) Differential Congo red staining: The effects of pH, non-aqueous solvents and the substrate.Histochem. J. 16, 195–210.
Millar, D. A. &Williams E. D. (1982) A step-wedge standard for the quantification of immunoperoxidase techniques.Histochem. J. 14, 609–20.
Nakanishi, I., Moriizuma, T., Ooi, A., Oda, Y. &Kajikawa, K. (1983) An ultrastructural study of periductal elastosis in human breast tumours.Acta Path. Jpn 33, 761–72.
Natali, B. G., Galloway, D., Nicotra, M. R. &De Martino, C. (1981) Topographic association of fibronectin with elastic fibres in the arterial wall. An immunohistochemical study.Conn. Tiss. Res. 8, 199–204.
Pearse, A. G. E. (1968)Histochemistry: Theoretical and Applied, Vol. 1 3rd edn. London: Churchill.
Ross, R. (1973) The elastic fibre: A review.J. Histochem. Cytochem. 21, 199–208.
Ross, R. &Bornstein, P. (1969) The elastic fibre. Separation and characterisation of its macromolecular components.J. Cell Biol. 40, 366–81.
Van Noorden, S. &Polak, J. M. (1983) Immunocytochemistry today, techniques and practice. InImmunocytochemistry (edited byPolak, J. M. andVan Noorden, S.), pp. 11–42. Bristol: John Wright & Sons.
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Mera, S.L., Lovett, F. & Davies, J.D. The effects of preliminary proteolysis on the immunohistochemical and dye staining properties of elastic fibres. Histochem J 17, 243–258 (1985). https://doi.org/10.1007/BF01003223
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DOI: https://doi.org/10.1007/BF01003223