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Determining Optimal Cytotoxic Activity of Human Her2neu Specific CD8 T cells by Comparing the Cr51 Release Assay to the xCELLigence System
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Determining Optimal Cytotoxic Activity of Human Her2neu Specific CD8 T cells by Comparing the Cr51 Release Assay to the xCELLigence System

Determining Optimal Cytotoxic Activity of Human Her2neu Specific CD8 T cells by Comparing the Cr51 Release Assay to the xCELLigence System

DOI:
10.3791/3683-v

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11:31 min

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August 08, 2012

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, ,
1Department of Immunology,College of Medicine, Mayo Clinic

Chapters

  • 00:05Title
  • 01:16Generation of Short-term Human Antigen-specific CD8 T Cell Lines
  • 02:55Preparation of Breast Cancer Cells
  • 04:19Co-culture of Tumor with CD8 T Cells
  • 05:45Chromium Release Assay (CRA)
  • 07:21HER-2/neu p369-specific T Cell / SKBR3 Breast Cancer Cells Co-cultures
  • 11:09Conclusion

Summary

Automatic Translation

Automatic Translation

The chromium release assay, a common assay for detecting cytotoxic T cell activity, has several limitations. Using antigen-specific CD8 T cells and the human breast cancer tumor line, SKBR3, in the present article, an impedance-based approach was examined for the capability of detecting cell killing.

Tags

Cytotoxic ActivityCD8 T CellsCr51 Release AssayXCELLigence SystemT Cell Receptor (TCR)Antigen-specific CD8 T CellsELIspotFlow CytometryCytotoxicity AssayChromium Release Assay (CRA)Gamma RadiationReproducibilityCaspase ReleaseBLT Esterase ActivityCD107 ExpressionImpedance-based AssayRoche XCelligence System

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