title: Immunohistochemical detection of the expression of pro-inflammatory cytokines by ovine pulmonary macrophages.
abstract: 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.
authors.affiliation: Departamento de Anatomía y Anatomía Patológica Comparadas, Edificio de Sanidad Animal, Facultad de Veterinaria, Universidad de Córdoba, Campus Universitario de Rabanales, 14014 Córdoba, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Carrasco
authors.forename: A
authors.initials: A
authors.lastname: Núñez
authors.forename: P J
authors.initials: PJ
authors.lastname: Sánchez-Cordón
authors.forename: M
authors.initials: M
authors.lastname: Pedrera
authors.forename: M
authors.initials: M
authors.lastname: Fernández de Marco
authors.forename: F J
authors.initials: FJ
authors.lastname: Salguero
authors.forename: J C
authors.initials: JC
authors.lastname: Gómez-Villamandos
mesh_terms: 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
chemical_list: D016207:Cytokines; D005404:Fixatives; D007375:Interleukin-1; D015850:Interleukin-6; D011136:Polysorbates; D014409:Tumor Necrosis Factor-alpha