title: Antigen specific killing assay using CFSE labeled target cells.
abstract: Carboxyfluorescein diacetate succinimidyl ester (CFSE) can be used to easily and quickly label a cell population of interest for in vivo investigation. This labeling has classically been used to study proliferation and migration. In the method presented here, we have shortened the timeline after adoptive transfer to look at survival and killing of epitope specific CFSE labeled target cells. The level of specific killing of a CD8 + T cell clone can indicate the quality of the response, as their quantity may be misleading. Specific CD8+ T cells can become functionally exhausted over time with a decline in cytokine production and killing. Also, certain CD8 + T cell clones may not kill as well as others with differing TCR specificities. For effective Cell Mediated Immunity (CMI), antigens must be identified that produce not only adequate numbers of responding T cells, but also functionally robust responding T cells. Here we assess the percent cell specific killing of two peptide specific T cell clones in BALB/c mice.
authors.affiliation: Pathology and Laboratory Medicine, University of Wisconsin-Madison, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Durward
authors.forename: Jerome
authors.initials: J
authors.lastname: Harms
authors.forename: Gary
authors.initials: G
authors.lastname: Splitter
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte / Q000032:analysis / Q000276:immunology*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D013388:Succinimides / Q000737:chemistry*
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides