Immunohistochemical detection of strain-specific major histocompatibility complex class I antigens in paraffin-embedded rat osteochondral tissue

An immunohistochemical method using formalin-fixed, paraffin wax-embedded sections is described for detecting strain-specific major histocompatibility complex class I antigens in knee-joint tissue from DA and Lewis strains of rat. The fixed osteochondral tissues were additionally decalcified in form...

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Bibliographic Details
Published in:The Histochemical journal 1993-02, Vol.25 (2), p.140-143
Main Authors: GURUSINGHE, C. J, HICKEY, M. J, HURLEY, J. V, O'BRIEN, B. M
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal
Antigens
Avidin
Biological and medical sciences
Biotin
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Histocompatibility antigens (hla, h-2 and other systems)
Histocompatibility Antigens Class I - analysis
Immunoenzyme Techniques
Immunohistochemistry
Knee Joint - immunology
Molecular immunology
Paraffin Embedding
Rats
Rats, Inbred Strains
Species Specificity
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Summary:An immunohistochemical method using formalin-fixed, paraffin wax-embedded sections is described for detecting strain-specific major histocompatibility complex class I antigens in knee-joint tissue from DA and Lewis strains of rat. The fixed osteochondral tissues were additionally decalcified in formic acid before processing for paraffin wax embedding. For immunohistochemistry, two monoclonal antibodies, one specific for DA class I allele RT1Aa and the other for Lewis class I allele RT1A1, were used together with the avidin-biotin immunoperoxidase procedure. It was necessary to use strain-specific normal rat serum as a diluent for the antibodies to suppress cross-strain recognition. DA-specific antibody stained positively only on DA rat sections, not on Lewis rat sections, and Lewis-specific antibody stained positively only on Lewis rat sections, and not on DA. Positive staining was localized in the bone marrow, osteochondral cells and endothelium. We propose that the use of a decalcification medium may have enhanced the immunoreactivity of the tissue. The method described can be used on sections of allografts from the two strains of rat to assess morphologically the extent of cellular replacement of the graft by the host's cells.
ISSN:0018-2214
1573-6865
DOI:10.1007/BF00157986