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.
1. Cover image - p. i 2. Table of Contents - p. iii 3. Series Page - p. iii 4. Copyright - p. iii 5. Contributors - p. iii 6. Chapter 1: Live-cell imaging for analysis of the NK cell immunological synapse - p. 1 7. Chapter 2: Analyzing the activity of the proteases granzyme B and caspase-8 inside living cells using fluorescence localization reporters - p. 13 8. Chapter 3: T cell activation and effector function in the human Jurkat T cell model - p. 25 9. Chapter 4: A tractable microscopy- and flow cytometry-based method to measure natural killer cell-mediated killing and infiltration of tumor spheroids - p. 43 10. Chapter 5: Kinetic, imaging based assay to measure NK cell cytotoxicity against adherent cells - p. 63 11. Chapter 6: Correlative light and electron microscopy to explore the lytic immunological synapse between natural killer cells and cancer cells - p. 93 12. Chapter 7: Metrics of 2D immunological synapses in human T cells via high-content confocal cell imaging - p. 107 13. Chapter 8: Kinetic measurements of human CD8+ T cell cytotoxic activity in a 384-well plate format - p. 121 14. Chapter 9: Imaging PIP2 and BCR microclusters in B cell immunological synapse - p. 135 15. Chapter 10: Activation kinetics of regulatory molecules during immunological synapse in T cells - p. 149 16. Chapter 11: Immunological synapse-driven transfer of extracellular vesicle microRNAs in primary lymphocytes - p. 173 17. Chapter 12: Generation of tumor spheroids in microwells to study NK cell cytotoxicity, infiltration and phenotype - p. 195
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.
1. Cover image - p. i 2. Table of Contents - p. iii 3. Series Page - p. iii 4. Copyright - p. iii 5. Contributors - p. iii 6. Chapter 1: Live-cell imaging for analysis of the NK cell immunological synapse - p. 1 7. Chapter 2: Analyzing the activity of the proteases granzyme B and caspase-8 inside living cells using fluorescence localization reporters - p. 13 8. Chapter 3: T cell activation and effector function in the human Jurkat T cell model - p. 25 9. Chapter 4: A tractable microscopy- and flow cytometry-based method to measure natural killer cell-mediated killing and infiltration of tumor spheroids - p. 43 10. Chapter 5: Kinetic, imaging based assay to measure NK cell cytotoxicity against adherent cells - p. 63 11. Chapter 6: Correlative light and electron microscopy to explore the lytic immunological synapse between natural killer cells and cancer cells - p. 93 12. Chapter 7: Metrics of 2D immunological synapses in human T cells via high-content confocal cell imaging - p. 107 13. Chapter 8: Kinetic measurements of human CD8+ T cell cytotoxic activity in a 384-well plate format - p. 121 14. Chapter 9: Imaging PIP2 and BCR microclusters in B cell immunological synapse - p. 135 15. Chapter 10: Activation kinetics of regulatory molecules during immunological synapse in T cells - p. 149 16. Chapter 11: Immunological synapse-driven transfer of extracellular vesicle microRNAs in primary lymphocytes - p. 173 17. Chapter 12: Generation of tumor spheroids in microwells to study NK cell cytotoxicity, infiltration and phenotype - p. 195
