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
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author Li, Tong
Smith, Mindy
Abdussamad, Maryam
Katz, Grace
Catalfamo, Marta
author_facet Li, Tong
Smith, Mindy
Abdussamad, Maryam
Katz, Grace
Catalfamo, Marta
author_sort Li, Tong
collection PubMed
description 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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spelling pubmed-97925532022-12-28 A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells Li, Tong Smith, Mindy Abdussamad, Maryam Katz, Grace Catalfamo, Marta STAR Protoc Protocol 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) Elsevier 2022-12-15 /pmc/articles/PMC9792553/ /pubmed/36527713 http://dx.doi.org/10.1016/j.xpro.2022.101939 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Li, Tong
Smith, Mindy
Abdussamad, Maryam
Katz, Grace
Catalfamo, Marta
A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title_full A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title_fullStr A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title_full_unstemmed A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title_short A flow-cytometry-based assay to assess granule exocytosis and GZB delivery by human CD8 T cells and NK cells
title_sort flow-cytometry-based assay to assess granule exocytosis and gzb delivery by human cd8 t cells and nk cells
topic Protocol
url 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
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