The Immunological Synapse Part A

1st Edition, Volume 173 - January 11, 2023
Imprint: Academic Press
Editors: Clément Thomas, Lorenzo Galluzzi
Language: English
Hardback ISBN:
9 7 8 - 0 - 3 2 3 - 9 0 1 5 4 - 3
eBook ISBN:
9 7 8 - 0 - 3 2 3 - 9 0 1 5 5 - 0

The Immunological Synapse, Part A, Volume 173 in the Methods in Cell Biology series provides state-of-the-art methods for the study of the immunological synapse. Sections cover Ima… Read more

Purchase options

BLOOM INTO SPRING SAVINGS

Save up to 25% on books and eBooks!

Shop now

Institutional subscription on ScienceDirect

Request a sales quote

The Immunological Synapse, Part A, Volume 173 in the Methods in Cell Biology series provides state-of-the-art methods for the study of the immunological synapse. Sections cover Imaging polarized granule release at the cytotoxic T cell immunological synapse using TIRF microscopy: control by polarity regulators, Analysis of centrosomal area actin reorganization and centrosome polarization upon lymphocyte activation at the immunological synapse, P815-based redirected degranulation assay to study human NK cell effector functions, Cytotoxic and Chemotactic Dynamics of Natural Killer Cells Quantified by Live-cell Imaging, Quantification of interaction frequency between antigen-presenting cells and T cells by conjugation assay, and more.

Other chapters focus on the Study of the Effects of NK-Tumor Cell Interaction by Proteomic Analysis and Imaging, Quantification of lymphocytic choriomeningitis virus specific T cells and LCMV viral titers, Quantification of lymphocytic choriomeningitis virus specific T cells and LCMV viral titers, An in vitro model to monitor natural killer cell effector functions against primary breast cancer, and Standardized Protocol for the Evaluation of Chimeric Antigen Receptor (CAR)-modified Cell Immunological Synapse Quality using the Glass-supported Planar L.

Cover image
Title page
Table of Contents
Copyright
Contributors
Chapter 1: Imaging polarized granule release at the cytotoxic T cell immunological synapse using TIRF microscopy: Control by polarity regulators
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
5: Notes
Acknowledgments
Declaration of interests
References
Chapter 2: Analysis of centrosomal area actin reorganization and centrosome polarization upon lymphocyte activation at the immunological synapse
Abstract
1: Introduction
2: Materials, cells, immunological synapse formation and image capture
3: Imaging the immunological synapse
4: Discussion, future perspectives and concluding remarks
5: Notes
Acknowledgments
Conflict of interest
Author contributions
References
Chapter 3: P815-based redirected degranulation assay to study human NK cell effector functions
Abstract
1: Introduction
2: Prior to the assay
3: Redirected degranulation assay
4: Flow cytometry staining
5: Analyzing CD16-induced effector functions of NK cells
6: Concluding remarks
7: Notes
Acknowledgments
Conflict of interests
References
Chapter 4: Cytotoxic and chemotactic dynamics of NK cells quantified by live-cell imaging
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
Acknowledgments
References
Chapter 5: Quantification of interaction frequency between antigen-presenting cells and T cells by conjugation assay
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
5: Notes
Acknowledgment
References
Chapter 6: Assessment of membrane lipid state at the natural killer cell immunological synapse
Abstract
1: Introduction
2: Materials
3: Common procedures
4: Assessment of membrane lipid state via confocal microscopy
5: Assessment of membrane lipid state via total internal reflection fluorescence microscopy
6: Data analysis
7: Notes
Acknowledgments
References
Chapter 7: Study of the effects of NK-tumor cell interaction by proteomic analysis and imaging
Abstract
1: Introduction
2: Melanoma/NK cell co-culture
3: Cell imaging
4: Analysis of the proteomic changes related to the process of EMT induced by NK cells
Notes
Acknowledgments
Declaration of interests
References
Chapter 8: Protocol for the murine antibody-dependent cellular phagocytosis assay
Abstract
1: Introduction
2: Materials
3: Notes
Acknowledgment
Author disclosure
References
Chapter 9: Quantification of lymphocytic choriomeningitis virus specific T cells and LCMV viral titers
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Conclusion
Acknowledgments
References
Chapter 10: An in vitro model to monitor natural killer cell effector functions against breast cancer cells derived from human tumor tissue
Abstract
1: Introduction
2: Before you begin
3: Key resources table
4: Materials and equipment
5: Step-by-step method details
6: Expected outcomes
7: Quantification and statistical analysis
8: Advantages
9: Limitations
10: Safety considerations and standards
References
Chapter 11: Standardized protocol for the evaluation of chimeric antigen receptor (CAR)-modified cell immunological synapse quality using the glass-supported planar lipid bilayer
Abstract
1: Introduction
2: Before you begin
3: Key resources table
4: Materials and equipment
5: Step-by-step method details
6: Expected outcomes
7: Statistical analysis
8: Limitations
References
Chapter 12: Potency monitoring of CAR T cells
Abstract
1: Introduction
2: Preparation of cells
3: Setup co-culture for extended long-term killing (ELTK) assay (Assay #1)
4: Setup co-culture for re-challenge assay (Assay #2)
5: General procedures of flow cytometry analysis
6: Analysis of CAR T cell killing and T cell counts by flow cytometry
7: Analysis of CAR T cell phenotypes by flow cytometry
8: Quantification of cytokine production by CAR T cells
9: Concluding remarks
10: 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

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health