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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.
  • Li T; Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA.
  • Smith M; Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20852, USA.
  • Abdussamad M; Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA.
  • Katz G; Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA.
  • Catalfamo M; Department of Microbiology and Immunology. Georgetown University School of Medicine, Washington, DC 20007, USA. Electronic address: mc2151@georgetown.edu.
STAR Protoc ; 4(1): 101939, 2023 03 17.
Article en En | MEDLINE | ID: mdl-36527713
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.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Células Asesinas Naturales / Linfocitos T CD8-positivos Límite: Humans Idioma: En Año: 2023 Tipo del documento: Article

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Células Asesinas Naturales / Linfocitos T CD8-positivos Límite: Humans Idioma: En Año: 2023 Tipo del documento: Article