Assessment of annexin A1 release during immunogenic cell death

Methods Enzymol. 2019:629:71-79. doi: 10.1016/bs.mie.2019.06.010. Epub 2019 Jul 2.

Abstract

The protein annexin A1 (ANXA1) belongs to the danger-associated molecular patterns (DAMPs) that alert the innate immune system about tissue perturbations. In the context of immunogenic cell death (ICD), ANXA1 is released from the cytoplasm of dying cells and, once extracellular, acts on formyl peptide receptor 1 (FPR1) expressed on dendritic cells to favor long-term interactions between dying and dendritic cells. As a result, the accumulation of extracellular ANXA1 constitutes one of the hallmarks of ICD. In the past, the detection of ANXA1 was based on semiquantitative immunoblots. More recently, a commercial enzyme-linked immunosorbent assay (ELISA) has been developed to measure ANXA1 in an accurate fashion. Here, we detail the protocol to measure the concentration of ANXA1 in the supernatants of cancer cells treated with chemotherapy.

Keywords: Annexin A1; Cancer; Chemotherapy; FPR1; Immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Annexin A1 / analysis*
  • Annexin A1 / immunology
  • Annexin A1 / metabolism
  • Antineoplastic Agents / pharmacology
  • Cell Line, Tumor
  • Cytoplasm / drug effects
  • Cytoplasm / immunology
  • Cytoplasm / metabolism
  • Drug Monitoring / instrumentation
  • Drug Monitoring / methods
  • Enzyme-Linked Immunosorbent Assay / instrumentation
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Extracellular Space / immunology
  • Extracellular Space / metabolism
  • Humans
  • Immunogenic Cell Death / drug effects*
  • Neoplasms / drug therapy*
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Reagent Kits, Diagnostic

Substances

  • ANXA1 protein, human
  • Annexin A1
  • Antineoplastic Agents
  • Reagent Kits, Diagnostic