Application Note: Quantitative Analysis of CAR-T Cell Exhaustion Using Advanced Flow Cytometry
T-cell exhaustion is a distinct phenotype associated with effector and memory T cells that can no longer perform their robust immune functions. It is triggered by chronic antigen stimulation, such as over expression of inhibitory receptors PD-1, CTLA-4, TIM-3, LAG-3 and CD244.
Exhaustion plays an important role in the tumor microenvironment (TME) where it inhibits the normal immune response against cancer. The same immune-inhibitory networks that influence endogenous T-cell function can also limit the efficacy of autologous T-cell therapy.
This application note describes how you can use the iQue® Advanced Flow Cytometry Platform and the iQue® Human T Cell Exhaustion Kit for studying T-cell exhaustion. We use this protocol to simultaneously quantify T-cell exhaustion markers and cytokines that are relevant to drug discovery. We also monitored the exhaustion status of CAR-T cells in response to antigen challenge, over time.
Key Protocol Benefits:
- Microvolume sampling, multiplexed interrogation and automated analysis simplify characterization and generate more insight from small sample volumes.
- Easy to use mix-and-read kits with optimized antibodies and cell health dyes allow you to capture both phenotypic and cytokine data simultaneously.
- Pre-designed gating strategy in conjunction with the intelligent iQue Forecyt® software enable easy analysis.
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