(89f) A Novel Experimental Approach to Investigate Cytokine Signaling Networks of Human T Cells

T cells are important for adaptive immune responses, and understanding the nature of their contributions are critical to identify immunological signatures for diseases, as well as define successful responses to therapeutic interventions. In recent years, it has become evident that the functional responses of differentiated T cells may be more dynamic than previously considered, but existing tools for characterizing activated human T cells (i.e., ELISpot and intracellular cytokine staining) cannot monitor the evolution of responses by individual cells or how their responses modulate each other. In this work, we present an application of microengraving to track the functional evolution and cytokine regulation of T cells. Our experimental platform evaluates over 10,000 individual cells in parallel, and generates up to 30 data points per cell that describe the cytokine secretion from eight distinct subsets of primary CD3+ T cells. Resulting dynamic measurements quantify the evolution of single-cell cytokine secretion following stimulation, offering a unique multi-dimensional approach to investigate the dynamics of specific immunophenotypes.  Using this technology, we explore the effect of different autocrine and paracrine signals on the dynamic secretion profiles of individual cells.  Such experimental analysis may offer insight on the extracellular cytokine networks as well as intracellular regulation of immune signaling pathways.