A Rapid High-Throughput Workflow For TCR Discovery On Solid Tumors With Functional Verification

T cell-mediated tumor death relies on complex mechanisms and cell-cell interactions, making the development of T cell-based therapies inherently challenging. Understanding T cell  differentiation and function is critical for teasing apart the complexity of the tumor microenvironment, however, T cells are exceedingly heterogeneous in both phenotype and function and have variable responses to stimulation. A variety of techniques are currently used to assess T cell function, including flow cytometry, live-cell imaging, and chromium release to assess target cell killing; and intracellular cytokine staining, ELISA, and ELISPOT to measure cytokine expression. However, as these techniques are necessarily applied to separate cell populations, any relationships can only be inferred, and multiple functional events can never be directly observed on the same cell. Perhaps more importantly, these typically are endpoint assays where the underlying processes are difficult to decipher longitudinally, and the recovery of live cells of interest for expansion and genotypic analysis is nearly impossible. In this study, we demonstrate the combination of a high-throughput platform that enables TCR discovery on solid tumors with direct functional annotation through our Opto™ Cell Therapy Development 1.0 workflow in combination with our Adherent Cell Kit. Furthermore, we are now able to evaluate cell products on a single-cell basis to identify and understand the direct relationships between the cell’s cytokine profile and its effector function subsets.