title: Assessment of Antigen-Specific Cellular Immunogenicity Using Intracellular Cytokine Staining, ELISpot, and Culture Supernatants.
abstract: Quantification of cytokine production by CD4 and CD8 T cells after in vitro recall stimulation with the immunizing antigen is a powerful approach to characterize the cellular immune responses to immunization. Here we describe three complementary methods for such quantification including flow cytometric analysis of cytokine production by intracellular staining, ELISpot determination of the numbers of cytokine-producing cells, and generation of secreted cytokines and chemokines in culture supernatants for analysis by ELISA and/or cytometric bead arrays.
authors.affiliation: Infectious Disease Research Institute, Seattle, WA, USA.
authors.forename: Elyse A
authors.initials: EA
authors.lastname: Beebe
authors.affiliation: Infectious Disease Research Institute, Seattle, WA, USA. mark.orr@idri.org.|Department of Global Health, University of Washington, 1616 Eastlake Ave E, Suite 400, Seattle, WA, USA. mark.orr@idri.org.
authors.forename: Mark T
authors.initials: MT
authors.lastname: Orr
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology* / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D017077:Culture Media, Conditioned / Q000737:chemistry; D016207:Cytokines / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D007111:Immunity, Cellular; D051379:Mice; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000941:Antigens; D017077:Culture Media, Conditioned; D016207:Cytokines
keywords: ELISA; ELISpot; Flow cytometry; Immunogenicity; Intracellular cytokine staining; T cell