Cytofluorometric assessment of dendritic cell-mediated uptake of cancer cell apoptotic bodies

Martina Musella; Gwenola Manic; Claudia Galassi; Ilio Vitale; Antonella Sistigu

doi:10.1016/bs.mie.2019.08.005

Cytofluorometric assessment of dendritic cell-mediated uptake of cancer cell apoptotic bodies

Methods Enzymol. 2020:632:39-54. doi: 10.1016/bs.mie.2019.08.005. Epub 2019 Oct 18.

Authors

Martina Musella¹, Gwenola Manic², Claudia Galassi³, Ilio Vitale⁴, Antonella Sistigu⁵

Affiliations

¹ Unit of Tumor Immunology and Immunotherapy, Department of Research, Advanced Diagnostics, and Technological Innovation, IRCCS Regina Elena National Cancer Institute, Rome, Italy; Department of Molecular Medicine, University "La Sapienza", Rome, Italy.
² Unit of Tumor Immunology and Immunotherapy, Department of Research, Advanced Diagnostics, and Technological Innovation, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
³ Istituto di Patologia Generale, Università Cattolica del Sacro Cuore, Rome, Italy.
⁴ Italian Institute for Genomic Medicine (IIGM), Turin, Italy; Candiolo Cancer Institute-FPO, IRCCS, Candiolo, Italy. Electronic address: iliovit@gmail.com.
⁵ Unit of Tumor Immunology and Immunotherapy, Department of Research, Advanced Diagnostics, and Technological Innovation, IRCCS Regina Elena National Cancer Institute, Rome, Italy; Istituto di Patologia Generale, Università Cattolica del Sacro Cuore, Rome, Italy. Electronic address: antonella.sistigu@gmail.com.

PMID: 32000907
DOI: 10.1016/bs.mie.2019.08.005

Abstract

Dendritic cells (DCs) are specialized antigen presenting cells (APCs) able to intake and crosspresent antigens (Ags) on major histocompatibility complex (MHC) class I and II molecules to T cells thus initiating primary and memory immune responses. DC-mediated Ag uptake and crosspresentation represent crucial steps toward cancer recognition and eventually elimination. Cytofluorometry is a standardized procedure to study phagocytosis. By fast and reproducible single cell measurements, flow cytometry allows for simultaneous biochemical and functional analyses of Ag intake. In this chapter, we discuss a two-color flow cytometric analysis of DC-mediated uptake of apoptotic bodies. We also show data on the adjuvanticity of Type-I-interferons (Type-I-IFNs) during Ag retention as we offer a guideline and a range of advice on sample preparation and acquisition.

Keywords: Antigen presenting cells; Cytofluorometry; Immunogenic cell death; Phagocytosis; Tumor.

Publication types

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

MeSH terms

Animals
Cell Line, Tumor
Coculture Techniques / methods
Dendritic Cells / immunology*
Extracellular Vesicles / immunology*
Flow Cytometry / methods*
Humans
Immunogenic Cell Death
Mice
Neoplasms / immunology*
Phagocytosis