Archives of Dermatological Research

, Volume 254, Issue 2, pp 113–120 | Cite as

Immunochemical localization ofin Vivo bound immunoglobulins in pemphigus vulgaris epidermis

Employment of a peroxidase-antiperoxidase multistep technique for light and electron microscopy
  • H. Hönigsmann
  • K. Holubar
  • K. Wolff
  • E. H. Beutner
Article

Summary

A multi-step immunocytochemical method utilizing horseradish peroxidase (HRP) as an immunological marker was employed to demonstrate, at the ultrastructural level, IgG antiepithelial autoantibodies boundin vivo to pemphigus epidermis, and circulating IgG antibodies, using monkey oesophagus as substrate. To demonstrate IgG the following antisera were employed in sequential steps: Goat antihuman IgG; rabbit antigoat IgG; goatanti-HRP-serum. After incubating with the final antigen, HRP, the latter was visualized with a cytochemical method. Results paralleled those obtained by immunofluorescence, the antiepithelial antibody being demonstrated at the sites of the intercellular spaces of the epidermis. Electron microscopic cytochemistry showed IgG in the surface coat of epidermal cells and in the intercellular space, both in the desmosomal and the interdesmosomal areas. These results confirm the results of a previous study but the localization of the antibody is more exact than that obtained with HRP-conjugates. The present method is more versatile and specific than HRP methods that utilize HRP-conjugated antibodies.

Keywords

Intercellular Space Pemphigus Final Antigen Cytochemical Method Monkey Oesophagus 

Zusammenfassung

Zum elektronenmikroskopischen Nachweisin vivo an Pemphigusepidermis gebundener antiepithelialer IgG-Autoantikörper und zirkulierender IgG-Antikörper wurde eine immuncytochemische Mehrstufenmethode mit Meerrettichperoxidase (HRP) verwendet. Folgende Antisera wurden zur Demonstration von IgG in aufeinander folgenden Schritten eingesetzt: Ziegen-antihuman-IgG, Kaninchen-anti-Ziegen-IgG und Ziegen-anti-HRP-Serum. Nach der Inkubation mit dem letzten Antigen, HRP, wurde dieses mittels einer cytochemischen Methode sichtbar gemacht. Die Ergebnisse stimmten mit denen der Immunfluorescenz völlig überein: Antiepitheliale Antikörper konnten im Intercellularraum der Epidermis aufgefunden werden. Die elektronenmikroskopisch-cytochemische Reaktion stellte IgG an der Oberfläche von Epidermalzellen und im Intercellularraum dar, und zwar sowohl in Desmosomen als auch im interdesmosomalen Bereich. Diese Ergebnisse bestätigen die Befunde einer früheren Untersuchung mit HRP-Konjugaten, die Lokalisation des Antikörpers ist jedoch exakter; die Methode ist vielseitiger und spezifischer als jene Methoden, bei welchen HRP konjugierte Antikörper verwendet werden.

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Copyright information

© Springer-Verlag 1975

Authors and Affiliations

  • H. Hönigsmann
    • 1
    • 2
  • K. Holubar
    • 1
    • 2
  • K. Wolff
    • 1
    • 2
  • E. H. Beutner
    • 1
    • 2
  1. 1.Division of Experimental Dermatology Department of Dermatology I.University of ViennaN. Y.USA
  2. 2.Vienna (Austria) and Department of MierobiologyState University of New York at BuffaloN. Y.USA

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