A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells

Abstract

Summary: CD8 T and NK cells mediate killing by delivery of perforin and granzyme B (GZB) stored in lysosome-like granules. We present a flow-cytometry-based protocol combined with a redirected killing assay to evaluate granule exocytosis and the cytotoxic potential of human CD8 T cells and NK cells. We describe the assessment of the delivered GZB inside the target cells. We then detail the detection of lysosome membrane protein CD107a exposed on the cell surface of the effector cells upon degranulation.For complete details on the use and execution of this protocol, please refer to Chen et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Flow Cytometry/Mass Cytometry
• Cell-based Assays
• Immunology