Modification of blood cell PrP epitope exposure during prion disease.
PrPC [normal cellular PrP (prion-related protein)] is a glycosylphosphatidylinositol-linked cell-surface glycoprotein that is expressed primarily by cells of the central and peripheral nervous system and the lymphoreticular system. During prion disease, PrPC undergoes structural modification to PrPSc (abnormal disease-specific conformation of PrP). The appearance of prion infectivity and PrPSc within different peripheral lymphoid tissue sites during natural scrapie infection in sheep is suggestive of haematogenic dissemination. For this to occur, blood cells may harbour or carry disease-associated PrP and in doing so present altered conformations of PrP on their cell-surface. In the present study, we show that changes in PrP epitope expression, or accessibility, can be detected on peripheral blood mononuclear cells during the course of experimental scrapie in susceptible sheep. Peripheral blood mononuclear cells isolated from VRQ homozygous lambs inoculated orally with scrapie were probed with either N- or C-terminal-specific anti-PrP monoclonal antibodies and analysed by flow cytometry. During the progression of scrapie, significant alterations were seen in the exposure of particular cell-surface PrP epitopes. These modifications included increased accessibility to N-terminal regions of the PrP molecule, to the region between beta-strand-2 and residue 171, and to the C-terminal region of helix-3. Increased accessibility in the globular C-terminal domain of PrP occurred in the vicinity of tyrosine dimers, which are believed to have increased solvent exposure in disease-associated PrP. We suggest that the alterations in anti-PrP monoclonal antibody recognition of cell-surface PrP on blood cells from scrapie-infected sheep are indicative of structural changes within this molecule that may be relevant to prion disease.
Thackray
Alana M
AM
Department of Veterinary Medicine, Centre for Veterinary Science, University of Cambridge, Madingley Road, Cambridge CB3 OES, UK.
Ryder
Stephen J
SJ
Bujdoso
Raymond
R
D000375:Aging / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D004912:Erythrocytes / Q000276:immunology*; D014644:Genetic Variation; D005838:Genotype; D018620:PrPSc Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D012608:Scrapie / Q000276:immunology*; D012756:Sheep / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D018620:PrPSc Proteins