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

Tong Li; Mindy Smith; Maryam Abdussamad; Grace Katz; Marta Catalfamo

doi:10.1016/j.xpro.2022.101939

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

STAR Protoc. 2023 Mar 17;4(1):101939. doi: 10.1016/j.xpro.2022.101939. Epub 2022 Dec 15.

Authors

Tong Li¹, Mindy Smith², Maryam Abdussamad¹, Grace Katz¹, Marta Catalfamo³

Affiliations

¹ Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA.
² Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20852, USA.
³ Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA. Electronic address: [email protected].

Abstract

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.

Keywords: Cell Biology; Cell-based Assays; Flow Cytometry/Mass Cytometry; Immunology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

CD8-Positive T-Lymphocytes* / metabolism
Cytoplasmic Granules / metabolism
Exocytosis
Granzymes / metabolism
Humans
Killer Cells, Natural*

Substances

Granzymes

Abstract

Publication types

MeSH terms

Substances

Grants and funding