Quantification of eIF2alpha phosphorylation during immunogenic cell death

Lucillia Bezu; Alejandra Wu Chuang; Juliette Humeau; Guido Kroemer; Oliver Kepp

doi:10.1016/bs.mie.2019.04.010

Quantification of eIF2alpha phosphorylation during immunogenic cell death

Methods Enzymol. 2019:629:53-69. doi: 10.1016/bs.mie.2019.04.010. Epub 2019 Apr 27.

Authors

Lucillia Bezu¹, Alejandra Wu Chuang², Juliette Humeau², Guido Kroemer³, Oliver Kepp⁴

Affiliations

¹ Service anesthésie-réanimation, Hôpital Européen Georges Pompidou, AP-HP, Paris, France; Equipe labellisée Ligue Nationale Contre le Cancer, Université Paris Descartes, Université Sorbonne Paris Cité, Université Paris Diderot, Institut National de la Santé et de la Recherche Médicale (INSERM), UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France.
² Equipe labellisée Ligue Nationale Contre le Cancer, Université Paris Descartes, Université Sorbonne Paris Cité, Université Paris Diderot, Institut National de la Santé et de la Recherche Médicale (INSERM), UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France; Faculty of Medicine, University of Paris Sud, Kremlin-Bicêtre, France.
³ Equipe labellisée Ligue Nationale Contre le Cancer, Université Paris Descartes, Université Sorbonne Paris Cité, Université Paris Diderot, Institut National de la Santé et de la Recherche Médicale (INSERM), UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France; Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France; Suzhou Institute for Systems Medicine, Chinese Academy of Medical Sciences, Suzhou, China; Department of Women's and Children's Health, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden. Electronic address: kroemer@orange.fr.
⁴ Equipe labellisée Ligue Nationale Contre le Cancer, Université Paris Descartes, Université Sorbonne Paris Cité, Université Paris Diderot, Institut National de la Santé et de la Recherche Médicale (INSERM), UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France. Electronic address: captain.olsen@gmail.com.

PMID: 31727256
DOI: 10.1016/bs.mie.2019.04.010

Abstract

Immunogenic cell death (ICD) is a particular modality of cell death that can be triggered by selected anticancer chemotherapeutics. Tumor cells undergoing ICD can induce an adaptive anticancer immune response that targets residual cancer cells with the same antigenic profile. The activation of a full-blown immune response against the tumor antigen is preceded by the release or exposure of danger associated molecular patterns (DAMPs) by tumor cells that stimulate the attraction, activation and maturation of dendritic cells and eventually the antigen-specific priming of cytotoxic T lymphocytes (CTLs). The phosphorylation of the eukaryotic translation initiation factor (EIF2A) is a pathognomonic characteristic of ICD, which governs the release/exposure of DAMPs such as ATP and calreticulin and thus the immunogenicity of cell death. Here we describe techniques to detect eIF2alpha phosphorylation for the assessment of ICD.

Keywords: Cancer; Endoplasmic reticulum stress; Immune response; Mitoxantrone; Oxaliplatin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alarmins / immunology
Alarmins / metabolism
Antigens, Neoplasm / immunology*
Antigens, Neoplasm / metabolism
Cell Culture Techniques / instrumentation
Cell Culture Techniques / methods
Cell Line, Tumor
Dendritic Cells / immunology
Dendritic Cells / metabolism
Doxorubicin / pharmacology
Epitopes, T-Lymphocyte / immunology
Epitopes, T-Lymphocyte / metabolism
Eukaryotic Initiation Factor-2 / immunology
Eukaryotic Initiation Factor-2 / isolation & purification*
Eukaryotic Initiation Factor-2 / metabolism
Fluorescent Antibody Technique, Direct / instrumentation
Fluorescent Antibody Technique, Direct / methods
Humans
Image Processing, Computer-Assisted / instrumentation
Image Processing, Computer-Assisted / methods
Immunogenic Cell Death / drug effects
Immunogenic Cell Death / immunology*
Microscopy, Fluorescence / instrumentation
Microscopy, Fluorescence / methods
Neoplasms / immunology*
Neoplasms / pathology
Oxaliplatin / pharmacology
Phosphorylation / immunology
Software
T-Lymphocytes, Cytotoxic / metabolism

Substances

Alarmins
Antigens, Neoplasm
Epitopes, T-Lymphocyte
Eukaryotic Initiation Factor-2
Oxaliplatin
Doxorubicin