Immunological Synapse

Numbered 173 in the Methods in Cell Biology series, this first part of The Immunological Synapse collects current laboratory procedures for studying the immunological synapse. Its opening chapters explain how to image polarized granule release at the cytotoxic T cell synapse using TIRF microscopy while controlling polarity regulators, and how to measure actin reorganization in the centrosomal area and centrosome polarization when lymphocytes are activated. Other methods include a P815-based redirected degranulation assay for examining the effector functions of human natural killer (NK) cells, live-cell imaging to quantify the cytotoxic and chemotactic behavior of NK cells, and a conjugation assay that measures how often antigen-presenting cells interact with T cells.

Further chapters turn to NK cell interactions with tumor cells, combining proteomic analysis with imaging. Others describe how to quantify lymphocytic choriomeningitis virus (LCMV)-specific T cells alongside LCMV viral titers, and set out an in vitro model for testing NK cell effector functions against primary breast cancer cells. The final chapter provides a standardized protocol for judging the quality of the immunological synapse formed by chimeric antigen receptor (CAR)-modified cells on a glass-supported planar surface.

Each method is laid out point by point, with accompanying notes on important considerations. The contributions come from an international group of prominent scientists working in the field.