Analysis of TCR/CD3 recycling at the immune synapse

Engagement of the T cell antigen receptor (TCR) by specific ligand bound to the major histocompatibility complex is the primary event that leads to the assembly of the immune synapse (IS). Central to this process is TCR clustering at the T cell-APC contact, which is achieved with the contribution of an endosomal pool that is delivered to the IS by polarized recycling. As the TCR recycling process has not been fully elucidated, we developed methods suitable to quantitate recycling to the plasma membrane of TCR/CD3 complexes that have been engaged at the cell surface and track their traffic through the intracellular vesicular compartments toward the IS.