Quantitative determination of phagocytosis by bone marrow-derived dendritic cells via imaging flow cytometry.

Abstract:

:Immunogenic cell death (ICD), induced by certain anticancer chemotherapeutics, leads to the emission of danger associated molecular patterns (DAMP) by cancer cells, which facilitates the attraction, activation and maturation of dendritic cells (DC) as well as the subsequent priming of effector T cells. In this context calreticulin (CALR) exposed at an early stage of ICD at the surface of the cancer cells serves as phagocytic signal and triggers the formation of immunological synapses between malignant cells and DC. Subsequent phagocytosis facilitates the transfer of tumor associated antigen and thus depicts a fundamental step in the generation of anticancer immunity. Here we provide an imaging flowcytometric protocol for the quantification of ICD-associated DC phagocytosis of cancer cells. As compared to the traditional flowcytometry-based analysis, the presented method offers additional means of differentiation between the transient formation of immunological synapses and the final DC-mediated phagocytosis of cancer cells.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Cerrato G,Liu P,Martins I,Kepp O,Kroemer G

doi

10.1016/bs.mie.2019.07.021

subject

Has Abstract

pub_date

2020-01-01 00:00:00

pages

27-37

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(19)30301-5

journal_volume

632

pub_type

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