Single-cell cytokine analysis to characterize CAR-T cell activation

Abstract

With the increase in the implementation of adoptive transfer anti-CD19 chimeric antigen receptor (CAR) T cell therapy, we introduce a novel platform to study their heterogeneous cytokine response at the single-cell level following activation. Here, we describe an activation platform which incubates single CAR-T cells with single-target CD19 antigen-presenting cells (APCs) and measures the resulting cytokine secretions. This can be compared to the more traditional bulk activation modalities. Both pipelines of activation are measured using our high-throughput, multiplexed single-cell secretomic microchip composed of an antibody barcode array and subnanoliter microchambers, capable of detecting up to 42 unique proteins. This platform has also been adapted to cell lines as well as primary cells.