IMMUNOHISTOCHEMICAL DETECTION OF PRION PROTEIN IN SHEEP WITH SCRAPIE

被引：90

作者：

MILLER, JM

JENNY, AL

TAYLOR, WD

MARSH, RF

RUBENSTEIN, R

RACE, RE

机构：

[1] Virology Cattle Research Unit, National Animal Disease Center, USDA, Agricultural Research Service, Ames, IA 50010

[2] National Veterinary Services Laboratories, USDA, Animal and Plant Health Inspection Service, Veterinary Services, Ames, IA 50010

[3] Department of Animal Health and Biomedical Sciences, University of Wisconsin, Madison

[4] Department of Virology, New York State Office of Mental Retardation and Developmental Disabilities, Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314

[5] Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIH, National Institute of Allergy and Infectious Diseases, Hamilton

来源：

JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION | 1993年 / 5卷 / 03期

关键词：

D O I：

10.1177/104063879300500301

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

0906 ;

摘要：

Prion protein (PrP), which is involved in the pathogenesis of scrapie, occurs in 2 forms. The form extracted from scrapie brain is protease resistant (PrP-res), whereas PrP from normal brain is protease sensitive (PrP-sen). This study examined whether PrP-res could be detected in brains of sheep with scrapie by immunohistochemistry (IHC). A suitable IHC procedure was developed using brain tissue from hamsters that had been inoculated with the transmissible mink encephalopathy agent. Tissue samples were fixed in PLP (periodate, lysine, paraformaldehyde) that contained paraformaldehyde at a concentration of 0.125%. Before application of the IHC technique, tissue sections were deparaffinized and treated with formic acid to simultaneously enhance PrP-res immunoreactivity and degrade PrP-sen. Primary antibody was obtained from a rabbit immunized to PrP-res extracted from brains of mice with experimentally induced scrapie. Brains from 21 sheep with histopathologically confirmed scrapie were examined by IHC. In all 21 brains, PrP-res was widely distributed throughout the brain stem. Staining was particularly intense in neuronal cell bodies and around blood vessels, The IHC technique successfully detected PrP-res in brain samples that had been frozen or that were severely autolyzed before fixation in PLP. Brains from 11 scrapie-suspect sheep that were not considered histologically positive were also examined by IHC. PrP-res was found in 4 of these brains. Sections of brains from 14 clinically normal sheep did not have detectable PrP-res. Results of this study indicate that IHC detection of PrP-res is equivalent, and perhaps superior, to histopathology for the diagnosis of scrapie in sheep. Furthermore, IHC is applicable to tissues that have autolytic changes or processing artifacts that prevent satisfactory histopathologic evaluation for lesions of scrapie.

引用

页码：309 / 316

页数：8

共 38 条

[1]

Bessen R.A., Marsh R.F., Identification of two biologically distinct strains of transmissible mink encephalopathy in hamsters, J Gen Virol, 73, pp. 329-334, (1992)

[2]

Brigati D.J., Budgeon L.R., Unger E.R., Immunocytochemistry is automated: Development of a robotic workstation based upon the capillary action principle, J Histotechnol, 11, pp. 165-183, (1988)

[3]

Brown P., Transmissible spongiform encephalopathies in humans: Kuru, Creutzfeldt-Jakob disease and Gerstmann-Straussler-Scheinker disease, Can J Vet Res, 54, pp. 38-41, (1990)

[4]

Bruce M.E., McBride P.A., Farquhar C.F., Precise targeting of the pathology of the sialoglycoprotein, PrP, and vacuolar degeneration in mouse scrapie, Neurosci Lett, 102, pp. 1-6, (1989)

[5]

Caughey B., Cellular metabolism of normal and scrapie-associated forms of PrP, Semin Virol, 2, pp. 189-196, (1991)

[6]

Chesebro B., Race R., Wehrly K., Identification of scrapie prion protein-specific mRNA in scrapie-infected and uninfected brain, Nature, 315, pp. 331-333, (1985)

[7]

Cunningham A.A., Bovine spongiform encephalopathy and British zoos, J Zoo Wildl Med, 22, pp. 304-308, (1991)

[8]

DeArmond S.J., Mobley W.C., DeMott D.L., Changes in the localization of brain prion proteins during scrapie infection, Neurology, 37, pp. 1271-1280, (1987)

[9]

Diedrich J.F., Bendheim P.E., Kim Y.S., Scrapie-associated prion protein accumulates in astrocytes during scrapie infection, Proc Natl Acad Sci USA, 88, pp. 375-379, (1991)

[10]

Farquhar C.F., Somerville R.A., Ritchie L.A., Post-mortem immunodiagnosis of scrapie and bovine spongiform encephalopathy, J Virol Methods, 24, pp. 215-222, (1989)

← 1 2 3 4 →