<em>In Vivo</em> Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells

Florian Stritzke; Hendrik Poeck; Simon Heidegger

doi:10.3791/62811

In vivo Triagem de imunogenicidade de vesículas extracelulares derivadas do tumor por ci...

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Cancer Research

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JoVE Journal Cancer Research

In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells

In vivo Triagem de imunogenicidade de vesículas extracelulares derivadas do tumor por citometria de fluxo de células T esplênicas

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08:02 min

September 23, 2021

DOI: 10.3791/62811-v

Florian Stritzke^1,2, Hendrik Poeck^1,2,3,4, Simon Heidegger^1,2

¹Department of Medicine III, School of Medicine,Technical University of Munich, ²Center for Translational Cancer Research (TranslaTUM), School of Medicine,Technical University of Munich, ³Department of Internal Medicine III,University Hospital Regensburg, ⁴National Centre for Tumor Diseases WERA

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Overview

This article demonstrates a method to assess the in vivo immunogenicity of tumor-derived extracellular vesicles (EVs) using flow cytometry. The protocol focuses on EVs from tumors undergoing treatment-induced immunogenic cell death, which are significant in tumor immunosurveillance.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Immunology

Background

Extracellular vesicles (EVs) play a role in intercellular communication.
EVs derived from tumors can indicate immunogenic responses.
Flow cytometry is a useful tool for analyzing EVs.
Understanding EVs can lead to personalized cancer treatments.

Purpose of Study

To assess the immunogenicity of tumor-derived EVs.
To demonstrate a flow cytometry method for EV analysis.
To explore the potential of EVs in cancer immunotherapy.

Methods Used

Preparation of cell culture media for EVs.
Ultracentrifugation to deplete bovine EVs from FCS media.
Flow cytometry for analyzing immunogenic properties of EVs.
Adaptation of the protocol for various experimental settings.

Main Results

The method effectively identifies immunogenic EVs.
Oxaliplatin-induced EVs showed significant immunostimulatory effects.
The protocol is adaptable for different cancer types.
Potential for personalized anti-cancer treatments using autologous EVs.

Conclusions

Flow cytometry is a valuable technique for EV analysis.
Understanding EV immunogenicity can enhance cancer therapies.
Further research is needed to explore clinical applications.

Frequently Asked Questions

What are extracellular vesicles?

Extracellular vesicles (EVs) are membrane-bound particles released by cells that play a role in cell communication and can carry proteins, lipids, and nucleic acids.

How does flow cytometry work?

Flow cytometry is a technique that uses lasers to analyze the physical and chemical characteristics of particles in a fluid as they pass through a detection apparatus.

Why are tumor-derived EVs important?

Tumor-derived EVs can provide insights into the tumor microenvironment and may serve as biomarkers for cancer diagnosis and treatment.

Can this method be used for other types of cancer?

Yes, the protocol can be adapted for various cancer types to assess the immunogenicity of tumor-derived EVs.

What is the significance of immunogenic cell death?

Immunogenic cell death triggers an immune response against tumor cells, which can enhance the effectiveness of cancer therapies.

How can EVs be used in personalized medicine?

Autologous EVs can be utilized to develop personalized anti-cancer treatments tailored to the individual patient's tumor characteristics.

Este manuscrito descreve como avaliar a imunogenicidade in vivo de vesículas extracelulares derivadas de células tumorais (EVs) usando citometria de fluxo. Os EVs derivados de tumores submetidos à morte de células imunogênicas induzidas pelo tratamento parecem particularmente relevantes na imunossuperveiência tumoral. Este protocolo exemplifica a avaliação dos EVs de tumor imunoestimulatório induzidos por oxaliplatina, mas pode ser adaptado a várias configurações.

Este vídeo mostra como avaliar a imunogenicidade in vivo de vesículas extracelulares derivadas do tumor, ou EVs, usando citometria de fluxo. O método de triagem descrito aqui é relativamente fácil de executar e pode ser adaptado a várias configurações. No câncer, identificando condições de liberação de vesícula extracelular imunogênica de particular relevância translacional.

Como tal, os EVs autólogos podem ser usados como tratamentos anticâncológicos personalizados. Demonstrando a primeira parte do procedimento estará Tatiana Nedelko, assistente técnica do meu laboratório. Para iniciar a preparação da mídia de cultura celular necessária para vesícula extracelular, ou EVs, esgotar os EVs bovinos em mídia FCS por ultracentrifugação a 100.000 vezes g por 24 horas a 4 graus Celsius.

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