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Evaluating Antibody-Dependent Cell-Mediated Cytotoxicity by Flow Cytometry

  • Irene van der Haar ÀvilaEmail author
  • Patricia Marmol
  • Jeannette Cany
  • Rolf Kiessling
  • Yago Pico de Coaña
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1913)

Abstract

Antibody-dependent cell-mediated cytotoxicity (ADCC) is a mechanism in which immune cell activation is induced by the cross-linking of CD16 with the Fc region of antibodies that at the same time bind specifically to cell surface antigens. ADCC stimulates the secretion of perforin, granzymes, and cytokines leading to lysis of the malignant cells. Natural killer (NK) cells express the CD16 receptor and can therefore be activated by ADCC to kill tumor cells. To study the cytotoxicity of NK cells against cancer cells, an ADCC-based assay is described: the flow cytometry-based cytotoxicity assay. In this method, the antibody trastuzumab, which binds specifically to HER2-positive malignant cells, is used to trigger ADCC.

Key words

Antibody cell-mediated cytotoxicity Natural killer cells Trastuzumab Flow cytometry 

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Copyright information

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

Authors and Affiliations

  • Irene van der Haar Àvila
    • 1
    • 2
    Email author
  • Patricia Marmol
    • 3
  • Jeannette Cany
    • 4
  • Rolf Kiessling
    • 1
  • Yago Pico de Coaña
    • 1
  1. 1.Department of Oncology-PathologyKarolinska InstituteStockholmSweden
  2. 2.Division of Tumor Biology and Immunology (B3)NKI-AvLAmsterdamThe Netherlands
  3. 3.Department of Cellular and Molecular BiologyKarolinska InstituteStockholmSweden
  4. 4.Laboratory Medicine, Department of HematologyRadboudumcNijmegenThe Netherlands

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