A protocol to study T-cell signaling in an immune synapse by microscopy

The immune synapse is a complex cellular structure that enables cell–cell communications between immune cells, mainly at the interface of an effector T-cell and an antigen-presenting cell (APC) that expresses the appropriate peptide–MHC complexes. With progressive technological advances, there has been increasing interest in understanding molecular events that take place in motile T-lymphocytes at the immune synapse. Here, we provide an optimized method to induce the formation of an immune synapse between a T-cell and an APC in vitro. The experimental protocol described herein would be useful in characterizing the role of cell surface molecules and downstream signaling pathways in the context of cell-to-cell communications between T-cells and APCs.