A Flow Cytometric NK Cell-Mediated Cytotoxicity Assay to Evaluate Anticancer Immune Responses In Vitro

  • Seila Lorenzo-Herrero
  • Christian Sordo-Bahamonde
  • Segundo González
  • Alejandro López-SotoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1884)


The immune system is able to detect and eliminate nascent and developing tumors. Thus, T lymphocytes of the adaptive immunity recognize cancer cells by detecting tumor-associated antigens, whereas certain innate immune cells scan for molecules that are mainly overexpressed on malignant and infected cells. The best example of the latter is natural killer (NK) cells. The activity of these immune cells is strictly governed by a balance between positive and negative signals provided by stress-regulated molecules that bind NK cell membrane receptors, such as the activatory receptor NKG2D. Given the key role of NK cells in eradicating cancer cells, the development of assays to study the cellular and molecular determinants of NK cell antitumor activity are of great relevance. In this chapter, we describe a flow cytometric in vitro assay to evaluate the cytotoxic activity of NK cells against a given target cell, including tumor cells. Moreover, this method is highly versatile, as it can be complemented with the use of antibodies blocking cell surface proteins (receptors or ligands) or a variety of chemical modulators, which allows the elucidation of molecules and signaling pathways that regulate NK cell anticancer function.

Key words

NK cells Cancer immunosurveillance NKG2D Blocking antibodies 



This work was supported by a Spanish grant from Instituto de Salud Carlos III (PI16/01485). S.L-H. holds a Severo Ochoa Fellowship by FICYT from Principado de Asturias, Spain (BP14–150).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Seila Lorenzo-Herrero
    • 1
    • 2
  • Christian Sordo-Bahamonde
    • 1
    • 2
  • Segundo González
    • 1
    • 2
  • Alejandro López-Soto
    • 3
    Email author
  1. 1.Departamento de Biología Funcional, InmunologíaUniversidad of OviedoOviedoSpain
  2. 2.Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Instituto de Investigación Sanitaria del Principado de Asturias (IISPA)OviedoSpain
  3. 3.Departamento de Biología Funcional, InmunologíaUniversidad de Oviedo, Instituto Universitario de Oncología del Principado de Asturias (IUOPA)OviedoSpain

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