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Related Experiment Video

Updated: Jun 6, 2026

Antigen Specific In Vivo Killing Assay using CFSE Labeled Target Cells
09:49

Antigen Specific In Vivo Killing Assay using CFSE Labeled Target Cells

Published on: November 9, 2010

Antigen specific killing assay using CFSE labeled target cells.

Marina Durward1, Jerome Harms, Gary Splitter

  • 1Pathology and Laboratory Medicine, University of Wisconsin-Madison, USA.

Journal of Visualized Experiments : Jove
|November 19, 2010
PubMed
Summary

This study introduces a faster method using Carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling to assess CD8+ T cell killing, revealing functional robustness over mere cell numbers for effective cell-mediated immunity.

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Area of Science:

  • Immunology
  • Cell Biology
  • In vivo studies

Background:

  • Carboxyfluorescein diacetate succinimidyl ester (CFSE) is a common tool for labeling cells in vivo.
  • Traditionally used for proliferation and migration studies, CFSE labeling can be adapted for other immunological investigations.

Purpose of the Study:

  • To evaluate the functional capacity of CD8+ T cell clones by assessing their killing ability.
  • To establish a shortened timeline for analyzing T cell-mediated killing post-adoptive transfer.
  • To determine if T cell quantity accurately reflects functional immune response quality.

Main Methods:

  • Adoptive transfer of epitope-specific CFSE-labeled target cells into BALB/c mice.
  • Shortened post-transfer observation period to assess cell survival and specific target cell killing.

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  • Quantification of specific CD8+ T cell-mediated killing of labeled target cells.

    • Main Results:

    • The study assessed the percentage of specific cell killing by two peptide-specific T cell clones.
    • Observed variations in killing efficiency among different T cell clones, highlighting TCR specificity impact.
    • Demonstrated that T cell killing assays provide a measure of functional response quality.

    Conclusions:

    • Assessing CD8+ T cell killing provides a more accurate measure of immune response quality than cell numbers alone.
    • Functional exhaustion and varying killing efficiencies of T cell clones underscore the importance of robust T cell responses.
    • This method allows for rapid evaluation of T cell effector function crucial for effective cell-mediated immunity.