Application Note: Characterization of T Cell Activation and Function in Cancer Immunotherapy using High-Throughput Cytometry

Authors: Veronica Bruce, Kirsty McBain and Nicola Bevan | Last updated: March 2024

Overview

T cells play critical roles in immunosurveillance and clearance of cells affected by viruses, bacteria and tumors. T cells have the ability to recognize and eliminate tumor cells. However, challenges such as poor tumor recognition and penetration into solid tumors, do hinder their efficacy.

Novel immunotherapies, including bispecific antibodies, tumor infiltrating lymphocytes (TILs), and chimeric antigen receptor (CAR) T cells, have been developed to enhance T cell responses against cancer.

There exists a need for development of robust and relevant strategies, to monitor novel therapeutics for sufficient activation and anti-tumor efficacy, while minimizing toxicity and off-target concerns.

To address the need for rapid monitoring of T cell function, we developed an optimized, high-content, multiplexed assay using high-throughput cytometry to measure T cell activation. This assay focuses on the use of the iQue® High-Throughput Screening (HTS) Cytometry Platform combined with the built-in, visual-based iQue Forecyt® software. The assay allows for assessment of multiparametric data of cell health, viability, phenotype, and effector function coupled with cytokine analysis from the same well, using simple workflows and minimal sample volumes.

This methodology also has the potential to predict on-target off-tumor effects and aid in the rapid identification of therapeutic candidates.


  • Document type: Application Note
  • Page count: 11
  • Read time: 15 minutes


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Application Note Sections include details of:

  • Antibody-Mediated T Cell Activation
  • CAR-T Cell Activation Assay
  • “On Target Off Tumor” CAR-T Profiling
  • Activation of Tumor Infiltrating Lymphocytes (TILs)

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