Methods for Analyzing Human T Cell Activation Through TCR or CAR Engagement

There are various ways to activate human T cells through their T Cell Receptor (TCR) or through overexpressed Chimeric Antigen Receptors (CARs), which are one of the most promising treatments in cancer immunotherapy. Here, we describe some basic methods to activate the human TCR or CAR using Antigen Presenting Cells presenting their natural antigen or superantigen, or stimulation with soluble antibodies or antibody-covered beads. There are several methods to analyze this activation and we describe here cytokine detection by ELISA, phosphoprotein detection by Western blot and expression of activation molecules at the cell surface by flow cytometry.