One of the issues impeding the application of chimeric antigen receptor T-cell (CAR-T) therapy is that some patients have T cell dysfunction, making treatment ineffective. To objectively screen these patients, we developed 3D fluorescence image analysis algorithm for quantifying cytotoxic behavior of CAR-T cells. Indicators include the area of immune synapses (IS), the actin depletion coefficient of IS, the actin migration rate, and the centrosome polarization angle. Our results demonstrate that these indicators are capable of differentiate CAR-T cell function.
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