Document Detail


Enhancement of immunohistochemical staining of scrapie proteins and immune cells within lymph nodes of early scrapie-infected sheep.
MedLine Citation:
PMID:  21722647     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Transmissible spongiform encephalopathies (TSE) are a group of fatal neurodegenerative diseases that affect animals as well as humans. The oldest of these diseases is Scrapie seen in sheep. Scrapie is caused by an altered form (PrP(sc)), capable of inducing "self-replication" of the normal host prion protein (PrP(c)). There is currently no universal standard for antigen retrieval when using immunohistochemistry to simultaneously stain the PrP(c) protein and other cellular markers. The use of formalin-fixed tissue creates a challenge by concealing the antigenic sites where an antibody would bind, and lengthy antigen retrieval methods must be applied in order to facilitate staining. Further complicating sheep tissue immunohistochemistry is a significant lack of commercial antibodies to sheep cell markers available in research. Here we developed a novel immunohistochemical technique using trypsin, formic acid, and hydrated autoclaving using citraconic anhydride buffer to increase sensitivity of staining for scrapie proteins and immune cell subsets. This allowed us to stain and identify cells within lymphoid tissue associated with early lymphoid pathogenesis in scrapie.
Authors:
Annissa Furr; David Knudsen; Michael B Hildreth; Alan J Young
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-06-23
Journal Detail:
Title:  Journal of immunological methods     Volume:  371     ISSN:  1872-7905     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-08-08     Completed Date:  2011-09-27     Revised Date:  2013-06-28    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1-7     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier B.V. All rights reserved.
Affiliation:
Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, United States.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal
Deer
Formates
Humans
Immunohistochemistry / methods,  veterinary*
Lymph Nodes / immunology,  metabolism,  pathology
PrPSc Proteins / immunology*,  metabolism*
Scrapie / immunology*,  metabolism*,  pathology
Sheep
Staining and Labeling / methods
Trypsin
Wasting Disease, Chronic / immunology,  metabolism,  pathology
Grant Support
ID/Acronym/Agency:
1R15AI072757/AI/NIAID NIH HHS; R15 AI072757-01/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Formates; 0/PrPSc Proteins; 64-18-6/formic acid; EC 3.4.21.4/Trypsin
Comments/Corrections

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