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Journal of Histochemistry and Cytochemistry, Vol. 50, 1357-1370, October 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Phenotyping of Protein–Prion (PrPsc)-accumulating Cells in Lymphoid and Neural Tissues of Naturally Scrapie-affected Sheep by Double-labeling Immunohistochemistry

Olivier Andréolettia, Patricia Berthonb, Etienne Levavasseurb, Daniel Marcb, Frédéric Lantierb, Eoin Monksc, Jean-Michel Elsend, and François Schelchera
a UMR INRA-ENVT, Physiopathologie Infectieuse et Parasitaire des Ruminants, Ecole Nationale Vétérinaire, Toulouse, France
b INRA, Laboratoire de Pathologie Infectieuse et Immunologie, Nouzilly, France
c Department of Agriculture, Food and Rural Development, Central Veterinary Research Laboratory, Abbotstown, Castleknock, Dublin, Ireland
d INRA, Station d'Amélioration Génétique des Animaux, Auzeville, France

Correspondence to: Olivier Andréoletti, Physiopathologie Infectieuse et Parasitaire des Ruminants, Ecole Nationale Vétérinaire, 23 Chemin des Capelles, 31076 Toulouse Cedex 3, France. E-mail: o.andreoletti@envt.fr

Transmissible spongiform encephalopathies are fatal neurodegenerative diseases characterized by amyloid deposition of protein–prion (PrPsc), the pathogenic isoform of the host cellular protein PrPc, in the immune and central nervous systems. In the absence of definitive data on the nature of the infectious agent, PrPsc immunohistochemistry (IHC) constitutes one of the main methodologies for pathogenesis studies of these diseases. In situ PrPsc immunolabeling requires formalin fixation and paraffin embedding of tissues, followed by post-embedding antigen retrieval steps such as formic acid and hydrated autoclaving treatments. These procedures result in poor cellular antigen preservation, precluding the phenotyping of cells involved in scrapie pathogenesis. Until now, PrPsc-positive cell phenotyping relied mainly on morphological criteria. To identify these cells under the PrPsc IHC conditions, a new, rapid, and highly sensitive PrPsc double-labeling technique was developed, using a panel of screened antibodies that allow specific labeling of most of the cell subsets and structures using paraffin-embedded lymphoid and neural tissues from sheep, leading to an accurate identification of ovine PrPsc-accumulating cells. This technique constitutes a useful tool for IHC investigation of scrapie pathogenesis and may be applicable to the study of other ovine infectious diseases.

(J Histochem Cytochem 50:1357–1370, 2002)

Key Words: PrPsc double labeling, immunohistochemistry, ovine scrapie


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