title: Detection of Avian Antigen-Specific T Cells Induced by Viral Vaccines.
abstract: Live attenuated viral vaccines are widely used in commercial poultry production, but the development of new effective inactivated/subunit vaccines is needed. Studies of avian antigen-specific T cells are primarily based on analyses ex vivo after activating the cells with recall antigen. There is a particular interest in developing robust high-throughput assays as chicken vaccine trials usually comprise many individuals. In many respects, the avian immune system differs from the mammalian, and T cell assessment protocols must be adjusted accordingly to account for, e.g., differences in leukocyte subsets.The carboxyfluorescein succinimidyl ester (CFSE) method described in this chapter has been adapted to chicken cells. In this test, cells of interest are stained with CFSE. The succinimidyl ester group covalently binds to cellular amines forming fluorescent conjugates that are retained in the cells even throughout division. This leads to daughter cells containing half the fluorescence of their parents. When lymphocytes are loaded with CFSE prior to ex vivo stimulation with specific antigen, the measurement of serial halving of its fluorescence by flow cytometry identifies the cells responding to the stimulation. This method has been successfully applied to studies of chicken antigen-specific T cells.
authors.affiliation: Department of Animal Science, Aarhus University, Tjele, Denmark. tina.dalgaard@anis.au.dk.
authors.forename: Tina Sørensen
authors.initials: TS
authors.lastname: Dalgaard
authors.affiliation: Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Liselotte Rothmann
authors.initials: LR
authors.lastname: Norup
authors.affiliation: Department of Animal Science, Aarhus University, Tjele, Denmark.
authors.forename: Helle Risdahl
authors.initials: HR
authors.lastname: Juul-Madsen
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D018929:Cell Culture Techniques; D002645:Chickens; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000378:metabolism; D010641:Phenotype; D013194:Staining and Labeling; D013388:Succinimides / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000956:Antigens, Viral; D005452:Fluoresceins; D013388:Succinimides; D014765:Viral Vaccines
keywords: Antigen-specific T cells; Carboxyfluorescein succinimidyl ester; Chicken viral vaccines; Flow cytometry; Proliferation; Vaccine-induced protective immunity