title: Generation of glycosylphosphatidylinositol linked chicken IL-17 to generate specific monoclonal antibodies applicable for intracellular cytokine staining.
abstract: Interleukin 17 (IL-17) cytokines play a crucial role in host defense and inflammatory diseases. Of the six mammalian IL-17 members five are described in the chicken (gg) genome. A novel method that attached cytokines to the surface of cells via a GPI linker was established to generate two chicken IL-17A and one chicken IL-17F specific mab. Recombinant gg IL-17A and gg IL-17F that showed dimerization in Western blot were used to verify the antibodies specificity. The mab could detect gg IL-17 by intracellular cytokine staining as demonstrated on cells expressing recombinant IL-17. Furthermore IL-17A and lower amounts of IL-17F were detectable in CD4 positive T cells of stimulated splenocytes. In conclusion, we have generated novel tools to analyze chicken IL-17 in more detail and demonstrated that the surface expression of cytokines is a reliable method to generate specific mab applicable for intracellular cytokine staining.
authors.affiliation: Institute for Animal Physiology, Department of Veterinary Sciences, University of Munich, Veterinärstrasse 13, 80539 Munich, Germany.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Walliser
authors.affiliation: Institute for Animal Physiology, Department of Veterinary Sciences, University of Munich, Veterinärstrasse 13, 80539 Munich, Germany. Electronic address: goebel@lmu.de.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Göbel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002645:Chickens / Q000276:immunology*; D017261:Glycosylphosphatidylinositols / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D020381:Interleukin-17 / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D017261:Glycosylphosphatidylinositols; D020381:Interleukin-17
keywords: Chicken; GPI-anchor; IL-17A; IL-17F; Intracellular cytokine staining