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Immunohistochemical identification of T and B lymphocytes in formalin-fixed paraffin-embedded chicken tissues using
1National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.
Insights
This study developed an immunohistochemical method to identify chicken T and B lymphocytes in formalin-fixed paraffin-embedded tissues. Commercial antibodies, particularly anti-human CD3 and anti-chicken BAFF-R, proved effective for avian disease diagnosis.
Area of Science:
- Veterinary Immunology
- Cellular Pathology
- Avian Pathology
Background:
- Immunohistochemistry (IHC) is crucial for diagnosing avian diseases and immunological analysis.
- Limited studies exist on IHC using commercial antibodies in formalin-fixed paraffin-embedded (FFPE) chicken tissues.
- Accurate identification of avian lymphocytes is essential for disease research.
Purpose of the Study:
- To establish a reliable immunohistochemical method for detecting chicken T and B lymphocytes in FFPE tissues.
- To evaluate the efficacy of commercially available antibodies for avian lymphocyte identification.
- To identify optimal antibodies for T and B lymphocyte detection in chickens.
Main Methods:
- Tested five commercial anti-T lymphocyte antibodies and ten commercial anti-B lymphocyte antibodies on FFPE chicken tissue sections.
- Utilized antibodies against chicken or human antigens for immunohistochemistry.
- Assessed antibody reactivity, specificity, and background staining.
Main Results:
- Five anti-T lymphocyte antibodies showed reactivity with chicken T lymphocytes.
- Three out of ten anti-B lymphocyte antibodies successfully detected chicken B lymphocytes.
- Anti-human CD3 (clone F7.2.38) and anti-chicken BAFF-R (clone 2C4) were identified as the most suitable antibodies.
- Optimal antibodies demonstrated specific staining in lymphoid tissues with minimal background.
Conclusions:
- A validated immunohistochemical method for detecting T and B lymphocytes in FFPE chicken tissues using commercial antibodies has been established.
- The identified antibodies provide reliable tools for avian disease diagnosis and immunological studies.
- This method enhances the capability for detailed analysis of avian immune responses.
Abstract:
Immunohistochemical method to detect avian lymphocytes is an efficient and reliable tool for accurate diagnosis, and immunological analysis of avian diseases. However, there are scarce studies reporting immunohistochemistry (IHC) using commercially available antibodies in formalin-fixed paraffin-embedded (FFPE) chicken tissues. In the present study, we established an immunohistochemical method to identify chicken T and B lymphocytes in FFPE chicken tissues using commercial antibodies against chicken or human antigens. For this IHC method, the five tested anti-T lymphocyte antibodies reacted with chicken T lymphocytes on the FFPE sections. Further, 10 commercial anti-B lymphocyte antibodies were tested; of these, three successfully detected chicken B lymphocytes for IHC. In particular, anti-human CD3 (clone F7.2.38) antibody was most suitable for the detection of chicken T lymphocytes, whereas anti-chicken B cell activating factor receptor (BAFF-R) antibody (clone 2C4) was most suitable for the detection of chicken B lymphocytes under our IHC staining conditions. These two antibodies reacted with numerous lymphocytes of all representative lymphoid tissues without problematic background staining and nonspecific reactions. Our results indicate that T and B lymphocytes in FFPE chicken tissues can be immunohistochemically detected using commercial antibodies.
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