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Updated: Jun 12, 2025
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Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
Published on: April 19, 2017
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Protocol for assessing murine cell doublet engagement and subsequent effects using flow cytometry and imaging flow
Yuval Shapir Itai1, Ziv Porat2, Rony Dahan1
1Department of Systems Immunology, Weizmann Institute of Science, Rehovot 7610001, Israel.
STAR Protocols
|September 22, 2024
Summary
This study introduces a new protocol to analyze immune cell interactions and bispecific engager effectiveness in tumors. It enables detailed assessment of cell-cell engagement and its impact on the tumor microenvironment.
Area of Science:
- Immunology
- Cancer Biology
- Biotechnology
Background:
- Physical interactions between immune and cancer cells are crucial in the tumor microenvironment.
- These interactions are key targets for novel immunotherapies like bispecific engagers.
Purpose of the Study:
- To present a protocol for evaluating murine cell doublet engagement using flow cytometry.
- To detail methods for identifying bispecific cell engager antibodies at cell interfaces.
- To quantify doublets and characterize cellular processes within them.
Main Methods:
- Utilizing flow cytometry and imaging flow cytometry for cell interaction analysis.
- Developing steps to identify bispecific antibodies at the cell-cell interface.
- Implementing doublet quantification and characterization of cellular morphology and processes.
Main Results:
- The protocol allows for the identification of bispecific cell engager antibodies.
- Quantitative analysis of cell doublets is achievable.
- Characterization of cellular protein morphology and processes within doublets is demonstrated.
Conclusions:
- The presented protocol provides a robust method for assessing bispecific cell engager activity in immune-mediated cellular interactions.
- This technique is valuable for understanding and developing immunotherapies targeting the tumor microenvironment.
