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Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses
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Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses

Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses

DOI:
10.3791/59531-v

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10:13 min

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May 06, 2019

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, ,
1Department of Microbiology and Immunology,Western University, 2Department of Medicine, Division of Clinical Immunology and Allergy,Western University, 3Department of Surgery, Division of General Surgery,Western University, 4Lawson Health Research Institute

Chapters

  • 00:04Title
  • 01:14T Antigen (Ag)-Expressing Tumor Cell Inoculation
  • 02:29Target Splenocyte Preparation
  • 04:10Target Splenocyte Coating with Irrelevant and Cognate Peptides and Carboxyfluorescein Succinimidyl Ester (CFSE) Labeling
  • 05:44CFSE-Labeling Evaluation
  • 06:50CFSE-Labelled Target Cell Injection into Nave and T-Ag-Primed Recipients
  • 07:31Data Acquisition
  • 08:21Results: Representative Analyses of CD8+ T Cell-Mediated Cytotoxicity
  • 09:36Conclusion

Summary

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We describe here a flow cytometry-based in vivo killing assay that enables examination of immunodominance in cytotoxic T lymphocyte (CTL) responses to a model tumor antigen. We provide examples of how this elegant assay may be employed for mechanistic studies and for drug efficacy testing.

Tags

In Vivo Cytotoxicity AssayTumor-specific CD8+ T Cell ResponsesImmunodominanceTumor AntigensCytotoxic T Cell ResponsesNK CellsInnate-like T LymphocytesAdherent Tumor Cell LineTarget Splenocyte PreparationC57BL/6 Mice

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