High throughput flow screening assays to profile cell-mediated killing

Traditional assays for monitoring cell-mediated killing are only capable of homogenous live/dead
readouts for an entire sample. IntelliCyt’s iQue Screener is a high throughput suspension screening
platform based on flow cytometry. The system can identify multiple cell types in solution and report on
multiple cell killing readouts, including cell viability and apoptosis. Here we demonstrate high
throughput flow assays for cell-mediated killing in examples using NK cells and chimeric antigen
receptor (CAR) T-cells. For the NK cell assay, NK92 cells were utilized as effector cells and Jurkat
cells were utilized as target cells. By labeling either target or effector cells with IntelliCyt’s MultiCyt FL4
Cell Proliferation and Encoder dye, both cell populations can be simultaneously monitored. Viability
was determined by cell membrane integrity and Caspase 3 activation for both Jurkat and NK92 cells,
and the specificity of the killing response was demonstrated using inhibitors of cell killing activity. In the
CAR T-cell assay, the efficacy of different CARs at targeting and killing a B-cell line (NALM-6) was
profiled with multiplex assays. In addition to cell encoding and live/dead analysis, cytokine responses
were evaluated using bead-based ELISA on the same analysis platform. These examples highlight the
robustness and flexibility of the iQue Screener for performing multiplexed screening assays for cell
mediated killing with greater contextual value.

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