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• W1549272413 abstract "This chapter describes an assay that helps bridge the gap between measurement of CD8+ T-cell cytolytic activity and the identity of the effector CD8+ T cells. Cytotoxic CD8+ T lymphocytes mediate the killing of target cells via two major pathways: (1) perforin-granzyme-mediated activation of apoptosis and (2) fas–fas ligand–mediated induction of apoptosis. Induction of these pathways depends on the release of cytolytic granules from the responding CD8+ T cells. Degranulation is a prerequisite to perforin-granzyme-mediated killing and is required for immediate lytic function mediated by responding antigen-specific CD8+ T cells. Cytotoxicity does not require de novo synthesis of proteins by the effector CD8+ T cell, instead, pre-formed lytic granules located within the cytoplasm are released in a polarized fashion toward the target cell. The lytic granules are membrane-bound secretory lysosomes containing a dense core composed of various proteins. The granule core is surrounded by a lipid bilayer containing numerous lysosomal-associated membrane glycoproteins (LAMPs). Thus, labeling responding cells with antibodies to CD107a and b and measuring their expression by flow cytometry can directly identify degranulating CD8+ T cells. The measurement of CD107 expression on the cell surface of activated T cells can be used with numerous other measures of T lymphocyte function. The chapter presents a protocol for this technique and then presents the results of the studies. Unlike standard intracellular cytokine assays, there are specific issues with examination of CD107a/b expression that must be considered during both the assay setup and the data analysis. The general applications for this technique revolve around the examination of antigen-specific T lymphocyte function in humans and mice." @default.
• W1549272413 created "2016-06-24" @default.
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• W1549272413 date "2004-01-01" @default.
• W1549272413 modified "2023-10-12" @default.
• W1549272413 title "Detection of T-Cell Degranulation: CD107a and b" @default.
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• W1549272413 doi "https://doi.org/10.1016/s0091-679x(04)75020-7" @default.
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