Protocol for assessing antibody-mediated cellular phagocytosis of lymphoma cells by human and murine macrophages in vitro

TL;DR

This paper provides a detailed in vitro protocol for measuring how macrophages engulf lymphoma cells using antibodies, which is important for cancer therapies.

Contribution

The paper introduces a standardized in vitro protocol for assessing antibody-dependent phagocytosis by macrophages from different sources.

Findings

The protocol outlines methods for generating mature macrophages from various sources.

It includes steps for evaluating the influence of agents on antibody-dependent phagocytosis.

The protocol helps identify the leading cell type responsible for changes in phagocytic activity.

Abstract

Antibody-dependent phagocytosis is an important process of cell engulfment and degradation by macrophages that holds therapeutical importance in the context of chemo-immunotherapy in, for example, lymphoma. Here, we present an in vitro protocol for determining the antibody-dependent phagocytosis of lymphoma cells by different types of macrophages under different treatment conditions. We describe steps for generating mature macrophages from different sources, usage of different clinically established antibodies, and elucidating the driving cell type of the observed changes in phagocytosis.

For complete details on the use and execution of this protocol, please refer to Beielstein et al.1

•Identify the antibody-dependent phagocytosis of macrophages from different origins•Determine the susceptibility toward phagocytosis of lymphoma cells•Evaluate the influence of agents on the antibody-dependent phagocytosis•Determine the leading cell type causing changed phagocytic activity

Identify the antibody-dependent phagocytosis of macrophages from different origins

Determine the susceptibility toward phagocytosis of lymphoma cells

Evaluate the influence of agents on the antibody-dependent phagocytosis

Determine the leading cell type causing changed phagocytic activity

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Antibody-dependent phagocytosis is an important process of cell engulfment and degradation by macrophages that holds therapeutical importance in the context of chemo-immunotherapy in, for example, lymphoma. Here, we present an in vitro protocol for determining the antibody-dependent phagocytosis of lymphoma cells by different types of macrophages under different treatment conditions. We describe steps for generating mature macrophages from different sources, usage of different clinically established antibodies, and elucidating the driving cell type of the observed changes in phagocytosis.

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