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Chimeric antigen receptor T cells (CAR T cells) therapy has revolutionarily changed the landscape of immunotherapy and been approved by the U. S Food and Drug Administration (FDA) since 2017 for several blood malignancies. To translate novel CAR T cells into clinical applications, it is essential to evaluate their antigen specificity, cytotoxic capacity, and off-target effects in vitro. A commonly used criteria to assess CAR T cell functionality involves detecting cytokine secretion following their engagement with target antigens. This chapter describes a method of combining intracellular cytokine staining and multi-color flow cytometry to measure CAR T cells activation following antigen stimulation.

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Measuring Chimeric Antigen Receptor T Cells (CAR T Cells) Activation by Coupling Intracellular Cytokine Staining with Flow Cytometry

  • 实验方案

  • 发表日期：

    2026年

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  • Chong Xu,
  • Yibo Yin

摘要

Chimeric antigen receptor T cells (CAR T cells) therapy has revolutionarily changed the landscape of immunotherapy and been approved by the U. S Food and Drug Administration (FDA) since 2017 for several blood malignancies. To translate novel CAR T cells into clinical applications, it is essential to evaluate their antigen specificity, cytotoxic capacity, and off-target effects in vitro. A commonly used criteria to assess CAR T cell functionality involves detecting cytokine secretion following their engagement with target antigens. This chapter describes a method of combining intracellular cytokine staining and multi-color flow cytometry to measure CAR T cells activation following antigen stimulation.