Evaluating Antibody-Dependent Cell-Mediated Cytotoxicity by Chromium Release Assay

Methods Mol Biol. 2019:1913:167-179. doi: 10.1007/978-1-4939-8979-9_12.

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 chromium release assay. In this method, the antibody trastuzumab, which binds specifically to HER2-positive malignant cells, is used to trigger ADCC.

Keywords: Antibody cell-mediated cytotoxicity; Chromium release assay; Natural killer cells; Trastuzumab.

MeSH terms

  • Antibody-Dependent Cell Cytotoxicity / drug effects
  • Antibody-Dependent Cell Cytotoxicity / immunology
  • Breast Neoplasms / blood
  • Breast Neoplasms / immunology
  • Breast Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Separation / instrumentation
  • Cell Separation / methods
  • Chromium Radioisotopes / metabolism*
  • Cytotoxicity Tests, Immunologic / instrumentation
  • Cytotoxicity Tests, Immunologic / methods*
  • Female
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods
  • GPI-Linked Proteins / immunology
  • GPI-Linked Proteins / metabolism
  • Humans
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / metabolism
  • Receptor, ErbB-2 / antagonists & inhibitors
  • Receptor, ErbB-2 / metabolism
  • Receptors, IgG / immunology
  • Receptors, IgG / metabolism
  • Trastuzumab / pharmacology

Substances

  • Chromium Radioisotopes
  • FCGR3B protein, human
  • GPI-Linked Proteins
  • Receptors, IgG
  • Chromium-51
  • ERBB2 protein, human
  • Receptor, ErbB-2
  • Trastuzumab