Category: Assay Development and Screening
Upon activation effector T cells undergo a profound metabolic reprogramming which allows the T cells to accommodate the energetic demands associated with rapid proliferation and production of copious amounts of inflammatory cytokines. This metabolic adaptation also leads to increased resistance of activated T cells to endogenous regulatory pathways leading to autoimmunity. Several lines of evidence suggest that targeting T cells metabolism could restore immune cell homeostasis by allowing endogenous regulatory pathways to regain control of pathogenic inflammatory T cells.
We successfully designed and executed a fully automated phosphoFlow phenotypic screen with the goal of identifying novel targets that can alter T cell metabolism. Both primary and secondary phenotypic assays were miniaturized and industrialized for this High Throughput Flow Cytometry assay going from a non-automated “one tube at-a-time” analysis format to a fully automated process in 384 well plate format, capable of analyzing more than 8000 samples per day. A combinatorial approach of proteomic profiling and screening with annotated small-molecules and siRNAs has revealed novel targets that have the potential to alter T cell metabolism without acute suppression of T cell effector functions.