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Immunology and Infection

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JoVE Journal Immunology and Infection
Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
 
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Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Article DOI: 10.3791/51105-v • 10:14 min • December 17th, 2013
December 17th, 2013 •
1, 1, 1
1Vaccine and Gene Therapy Institute of Florida

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This protocol describes a sensitive, cell-based cytotoxicity assay. By enumerating the decrease in frequency of live target CD4+ T cells in the presence of an increasing number of effector CD8+ T cells, this assay allows for the direct assessment of cytolytic activity of antigen-specific CD8+ T cells.

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Cell-based Flow Cytometry Assay Measure Cytotoxic Activity CD8+ T Cells Antigen-specific Kill CD4+ T Cells Cognate Peptide CFSE Pulsed Un-pulsed Mixed Incubated Purified CD8+ T Cells Autologous Target CD4+ T Cells Flow Cytometry Analysis Coculture Effector CD8+ T Cells Peptide/MHCI Tetramer Staining Specific Lysis Effector Versus Target Ratios Lytic Units LU30/106 Cells
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