In vitro and ex vivo evaluation of tumor-derived exosome-induced dendritic cell dysfunction in mouse

Exosomes that contain various signaling molecules, such as proteins, nucleotides, metabolites, and lipids, are important for intercellular communication. Dendritic cells (DC) are central regulators of anti-tumor immunity but can be suppressed by tumor-derived exosomes (TDEs) in the tumor microenvironment. Here, we describe a step-by-step protocol for TDE isolation and evaluation of TDEs on DCs both in vitro and ex vivo with high repeatability. This approach is useful for the interrogating TDE-DC interactions and identification of novel immune regulators. For complete details on the use and execution of this protocol, please refer to Yin et al. (2020). [Display omitted] •Separation of tumor-derived exosomes (TDEs) from tumor cell supernatant•Generation of TDE-treated DCs in vitro and ex vivo•Magnetic purification of CD8+ T cell from OT I transgenic mice spleen•Analysis of T cell proliferation via flow cytometry Exosomes that contain various signaling molecules, such as proteins, nucleotides, metabolites, and lipids, are important for intercellular communication. Dendritic cells (DC) are central regulators of anti-tumor immunity but can be suppressed by tumor-derived exosomes (TDEs) in the tumor microenvironment. Here, we describe a step-by-step protocol for TDE isolation and evaluation of TDEs on DCs both in vitro and ex vivo with high repeatability. This approach is useful for the interrogating TDE-DC interactions and identification of novel immune regulators.