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Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
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Measurement of polyclonal and antigen-specific cytotoxic T cell function

W E Biddison1, R Lichtenfels, M Adibzadeh

  • 1National Institute of Neurological Disorders and Stroke, Bethesda, Maryland, USA.

Current Protocols in Immunology
|April 25, 2008
PubMed

Measurement of in vitro cytotoxic function of human T cells can be accomplished by polyclonal stimulation of T cell effectors using anti-CD3 antibody, which stimulates all cytolytic effector cells, or with a specific stimulating antigen. Accordingly, two sets of assays of cytolytic T cell function are described in this unit, one for measuring anti-CD3-mediated cytotoxicity and the other for measuring antigen-specific cytotoxicity. Although the calcein release assay (CARE-LASS) described here is for use with antigen-activated cytotoxic T lymphocytes (CTL) as well as natural killer (NK) or lymphokine-activated killer (LAK) cells, minor changes in the protocols that address polyclonal T cell activation are described that make them suitable for use with calcein-labeled target cells.

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