The Immunological Synapse - Part B

1st Edition, Volume 178 - July 11, 2023
Editors: Clément Thomas, Lorenzo Galluzzi
Language: English
Hardback ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 1 6 3 - 3
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 1 6 4 - 0

The Immunological Synapse - Part B, Volume 178 in the Methods in Cell Biology series provides state-of-the-art methods for the study of the immunological synapse. This first vo… Read more

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The Immunological Synapse - Part B, Volume 178 in the Methods in Cell Biology series provides state-of-the-art methods for the study of the immunological synapse. This first volume covers various aspects on T cell and natural killer (NK) cell synapses, including imaging polarized granule release using TIRF microscopy, analysis of actin reorganization and centrosome polarization, redirected degranulation, live cell-imaging to quantify cytotoxic and chemotactic dynamics, quantification of interactions between APCs and T cells, assessment of membrane lipid state at the immunological synapse, proteomic analysis and imaging of NK-tumor cell interaction, evaluating natural killer cell effector functions against breast cancer cells derived from human tumor tissue, evaluation of chimeric antigen receptor (CAR)-modified cell immunological synapse quality using the glass-supported planar lipid bilayer, and monitoring potency of therapeutic CAR T cells. It also includes one chapter on quantification of lymphocytic choriomeningitis virus specific T cells and LCMV viral titers, and one chapter on the murine antibody-dependent cellular phagocytosis assay.

Cover image
Title page
Table of Contents
Series Page
Copyright
Contributors
Chapter 1: Live-cell imaging for analysis of the NK cell immunological synapse
Abstract
1: Introduction
2: Materials
3: Methods
Acknowledgments
References
Chapter 2: Analyzing the activity of the proteases granzyme B and caspase-8 inside living cells using fluorescence localization reporters
Abstract
1: Introduction
2: Material and methods
3: Concluding remarks
4: Notes
Acknowledgments
Declaration of interests
References
Chapter 3: T cell activation and effector function in the human Jurkat T cell model
Abstract
1: Introduction
2: Materials
3: Methods
4: Discussion
5: Notes
Acknowledgments
References
Chapter 4: A tractable microscopy- and flow cytometry-based method to measure natural killer cell-mediated killing and infiltration of tumor spheroids
Abstract
1: Introduction
2: Protocol
3: Notes
4: Concluding remarks
Acknowledgments
References
Chapter 5: Kinetic, imaging based assay to measure NK cell cytotoxicity against adherent cells
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
5: Notes
Acknowledgments
Declaration of interests
References
Chapter 6: Correlative light and electron microscopy to explore the lytic immunological synapse between natural killer cells and cancer cells
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
5: Notes
Acknowledgments
References
Chapter 7: Metrics of 2D immunological synapses in human T cells via high-content confocal cell imaging
Abstract
1: Introduction
2: Material
3: Procedure
4: Concluding remarks
5: Notes
Acknowledgments
References
Chapter 8: Kinetic measurements of human CD8+ T cell cytotoxic activity in a 384-well plate format
Abstract
1: Introduction
2: Material
3: Procedure
Acknowledgments
References
Chapter 9: Imaging PIP2 and BCR microclusters in B cell immunological synapse
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
Notes
Acknowledgments
Disclosure
References
Chapter 10: Activation kinetics of regulatory molecules during immunological synapse in T cells
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
Acknowledgments
Discussion
References
Chapter 11: Immunological synapse-driven transfer of extracellular vesicle microRNAs in primary lymphocytes
Abstract
1: Introduction
2: Material
3: Methods
4: Concluding remarks
5: Notes
Acknowledgments
References
Chapter 12: Generation of tumor spheroids in microwells to study NK cell cytotoxicity, infiltration and phenotype
Abstract
1: Introduction
2: Materials
3: Protocol
4: Notes
References

Clément Thomas

Clément Thomas, Ph.D. is a research group leader at the Department of Cancer Research of the Luxembourg Institute of Health, Luxembourg. He received his Ph.D. in Molecular and Cellular Biology from the University of Strasbourg. Over the last 15 years, his group has developed extensive knowledge and expertise in the mechanisms regulating actin cytoskeleton organization and dynamics, as well as in live cell imaging approaches. His current research is centered on the role of the actin cytoskeleton in tumor immune evasion.

Affiliations and expertise

Group Leader, Cytoskeleton and Cancer Progression, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg

Lorenzo Galluzzi

Lorenzo Galluzzi is Assistant Professor of Cell Biology in Radiation Oncology at the Department of Radiation Oncology of the Weill Cornell Medical College, Honorary Assistant Professor Adjunct with the Department of Dermatology of the Yale School of Medicine, Honorary Associate Professor with the Faculty of Medicine of the University of Paris, and Faculty Member with the Graduate School of Biomedical Sciences and Biotechnology of the University of Ferrara, the Graduate School of Pharmacological Sciences of the University of Padova, and the Graduate School of Network Oncology and Precision Medicine of the University of Rome “La Sapienza”. Moreover, he is Associate Director of the European Academy for Tumor Immunology and Founding Member of the European Research Institute for Integrated Cellular Pathology. Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. He has published over 450 articles in international peer-reviewed journals and is the Editor-in-Chief of four journals: OncoImmunology (which he co-founded in 2011), International Review of Cell and Molecular Biology, Methods in Cell biology, and Molecular and Cellular Oncology (which he co-founded in 2013). Additionally, he serves as Founding Editor for Microbial Cell and Cell Stress, and Associate Editor for Cell Death and Disease, Pharmacological Research and iScience.

Affiliations and expertise

Assistant Professor of Cell Biology in Radiation Oncology, Department of Radiation Oncology, Weill Cornell Medical College, NY, USA