Immunohistochemical detection of the expression of pro-inflammatory cytokines by ovine pulmonary macrophages.
The aim of this study was to determine the expression of three proinflammatory cytokines by pulmonary macrophages of sheep in paraffin wax-embedded tissue. Samples of lung from seven healthy sheep were fixed by immersion in either 10% neutral buffered formalin, acetic formalin, paraformaldehyde-lysine-periodate or Bouin's solution and processed for structural and immunohistochemical studies. The expression of interleukin (IL)-1alpha, IL-6 and tumour necrosis factor (TNF)-alpha by pulmonary intravascular macrophages (PIMs) and alveolar macrophages (AMs) was detected by the avidin-biotin-peroxidase (ABC) technique. Bouin's solution proved to be the most suitable fixative and Tween 20 the most effective pretreatment for increasing permeability. Constitutive expression of IL-1alpha, IL-6 and TNF-alpha by both macrophage populations was detected. The number of PIMs expressing IL-1alpha (the predominant cytokine in ovine lung) was higher than that of AMs, while the expression of IL-6 was greater in AMs. No differences between PIMS and AMs were found in respect of TNF-alpha expression. The evaluation of cytokine expression represents a valuable tool for studying the pathogenesis of disease in the ovine lung.
Carrasco
L
L
Departamento de Anatomía y Anatomía Patológica Comparadas, Edificio de Sanidad Animal, Facultad de Veterinaria, Universidad de Córdoba, Campus Universitario de Rabanales, 14014 Córdoba, Spain.
Núñez
A
A
Sánchez-Cordón
P J
PJ
Pedrera
M
M
Fernández de Marco
M
M
Salguero
F J
FJ
Gómez-Villamandos
J C
JC
D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects; D016207:Cytokines / Q000276:immunology*; D005404:Fixatives; D007124:Immunoenzyme Techniques / Q000662:veterinary*; D007375:Interleukin-1 / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D016676:Macrophages, Alveolar / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D011136:Polysorbates / Q000494:pharmacology; D012756:Sheep / Q000502:physiology*; D016707:Tissue Fixation / Q000662:veterinary; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D016207:Cytokines; D005404:Fixatives; D007375:Interleukin-1; D015850:Interleukin-6; D011136:Polysorbates; D014409:Tumor Necrosis Factor-alpha