Related Experiment Video
Updated: Jun 4, 2025
11:31
Determining Optimal Cytotoxic Activity of Human Her2neu Specific CD8 T cells by Comparing the Cr51 Release Assay to the xCELLigence System
Published on: August 8, 2012
25.0K
Protocol for assessing T cell receptor-mediated human T cell cytotoxicity.
Kathryne E Marks1, Ifeoluwakiisi Adejoorin2, John Sowerby1
1Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
STAR Protocols
|January 5, 2025
Summary
This study details a new protocol for measuring human T-cell cytotoxicity. The method assesses T-cell receptor (TCR) triggering and target cell killing using flow cytometry, applicable to small patient samples.
Area of Science:
- Immunology
- Cellular Biology
Background:
- Assessing human T-cell cytotoxicity is crucial but technically demanding.
- Existing methods may require large cell numbers, limiting clinical applications.
Purpose of the Study:
- To present a standardized protocol for measuring polyclonal human T-cell cytotoxicity.
- To enable T-cell killing assays using limited effector cell populations, such as those from patient samples.
Main Methods:
- Preparation of effector (T-cells) and target cells.
- Incubation of target cells with effector cells following T-cell receptor (TCR) triggering.
- Quantification of target cell death via flow cytometry.
Main Results:
- The protocol effectively measures T-cell mediated cytotoxicity.
- It is applicable to small numbers of effector T-cells, including those from clinical specimens.
- The assay relies on TCR triggering to initiate the cytotoxic response.
Conclusions:
- This protocol provides a robust and accessible method for assessing human T-cell cytotoxicity.
- It facilitates research and clinical applications involving T-cell function, particularly with limited cell availability.
