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Cytokine expression in paraffin wax-embedded tissues from conventional calves
M Pedrera1, P J Sánchez-Cordón, J L Romero-Trevejo
1Departamento de Anatomía y Anatomía Patológica Comparadas, Facultad de Veterinaria, Universidad de Córdoba, Edificio de Sanidad Animal, Campus de Rabanales, 14014, Córdoba, Spain.
Insights
This study identified key cytokine-producing cells in healthy calves, primarily macrophage-like cells expressing tumour necrosis factor-alpha (TNFα) and interleukins (IL). These findings are crucial for understanding calf inflammatory diseases.
Area of Science:
- Veterinary immunology
- Cellular biology
- Immunohistochemistry
Background:
- Cytokines like TNFα, IL-1, and IL-6 play critical roles in immune responses and inflammation.
- Understanding cytokine expression in healthy animals is fundamental for diagnosing and treating diseases.
Purpose of the Study:
- To evaluate antibody cross-reactivity against human and porcine cytokines (TNFα, IL-1α, IL-1β, IL-6).
- To determine the distribution of cytokine-expressing cells in healthy calf tissues.
- To optimize immunohistochemical techniques for cytokine detection in bovine tissues.
Main Methods:
- Immunohistochemistry using the avidin-biotin-peroxidase technique on paraffin-embedded calf tissues.
- Comparison of formalin and Bouin's solution as fixatives.
- Evaluation of Tween 20 for antigen retrieval to enhance detectability.
Main Results:
- Bouin's solution and Tween 20 were optimal for antigen unmasking and detection.
- Constitutive expression of TNFα, IL-1α, IL-1β, and IL-6 was detected, mainly in macrophage-like cells.
- Lymphoid organs showed higher immunolabelling than lung, liver, or kidney, with TNFα and IL-1α predominant in gut-associated lymphoid tissue and mesenteric lymph nodes.
Conclusions:
- Established immunohistochemical methods for detecting key cytokines in calves.
- Identified specific cell populations and tissues with high cytokine expression.
- Provides a foundation for future research into calf inflammatory conditions.
Abstract:
The cross-reactivity of antibodies against human tumour necrosis factor (TNF)alpha, interleukin (IL)-1alpha, IL-1beta and porcine IL-6, and the distribution of immunolabelled cells were evaluated on paraffin wax-embedded tissues from five healthy calves. The tissues were fixed in 10% buffered formalin or Bouin's solution and processed for structural studies and immunohistochemical studies by the avidin-biotin-peroxidase technique. Bouin's solution proved to be the more suitable fixative and Tween 20 the most effective antigen unmasking technique for increasing detectable antigenicity. Constitutive expression of TNFalpha, IL-1alpha, IL-1beta and IL-6 by different cell populations, mainly macrophage-like cells, was detected. Lymphoid organs displayed a higher presence of immunolabelled cells than did lung, liver or kidney. TNFalpha and IL-1alpha appeared as the predominant cytokines, especially in the gut-associated lymphoid tissue of the ileum and in the regional mesenteric lymph nodes. The results will facilitate investigation of the role of these cytokine-producing cells in inflammatory disease processes in calves.
