Determining Activation Status of Natural Killer Cells Following Stimulation via Cytokines and Surface Receptors

Methods Mol Biol. 2022:2463:181-194. doi: 10.1007/978-1-0716-2160-8_13.

Abstract

Stimulation of Natural Killer (NK) cells with cytokines, target cell interaction, or antibody mediated activation of receptors on the NK cell surface enables the dissection of specific signaling intermediates in different activation pathways. NK cell activation status is commonly measured by production of interferon gamma (IFNγ) and expression of the degranulation marker LAMP-1 (CD107a). Cytotoxic potency can also be assessed by the production of perforin, granzymes, and tumor necrosis factor alpha (TNFα). NK cell receptor mediated activation by antibodies requires crosslinking of the receptor-specific antibodies; thus, in vitro activation assays are performed by binding antibodies to cell culture plates. All parameters can be measured by flow cytometry.

Keywords: Activating receptors; Antibody-directed activation; Cytokine; Effector cell; Flow cytometry; IFNγCD107a; NK cell; Natural Killer cell; Target cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytokines* / metabolism
  • Interferon-gamma / metabolism
  • Killer Cells, Natural*
  • Lymphocyte Activation
  • Perforin / metabolism

Substances

  • Cytokines
  • Perforin
  • Interferon-gamma