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

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 descr...

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Detalles Bibliográficos
Autores principales: Li, Tong, Smith, Mindy, Abdussamad, Maryam, Katz, Grace, Catalfamo, Marta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9792553/
https://www.ncbi.nlm.nih.gov/pubmed/36527713
http://dx.doi.org/10.1016/j.xpro.2022.101939
Descripción
Sumario: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)

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