T-Cell Motility

This volume discusses the latest developments in cellular, molecular, biochemical, and imaging assays to study the biology and functions of T-cells. The chapters in this book cover topics such as LFA-1/ICAM-1 interactions in T-cell motility; using 3D-SIM to dissect signaling cross-talks in motile T-cells; GapmeR-mediated gene silencing in motile T-cells; activity of cellular kinases in migrating T-cells; and computational analysis of protein-protein interactions in motile T-cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, T-Cell Motility: Methods and Protocols is an essential resource for graduate students, postdoctoral fellows, and principal investigators working in the fields ofimmunology, T-cell biology, biochemistry, molecular biology, and imaging.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt
An Introduction to LFA-1/ICAM-1 Interactions in T-Cell Motility.- Isolation of Human Peripheral Blood T Lymphocytes.- A Laboratory Model to Study T-Cell Motility.- Quantification of T-Cell Migratory Phenotypes using High Content Analysis.- Live Cell Imaging and Analysis to Capture T-Cell Motility in Real-Time.- Three-Dimensional Structured Illumination Microscopy (3D-SIM) to Dissect Signalling Cross-Talks in Motile T-Cells.- Real-Time Impedance-Based Detection of LFA-1-Stimulated T-Cell Transwell Chemotaxis.- Quantitative Real-Time PCR for Evaluating Transcriptional Changes in T-Lymphocytes.- GapmeR-Mediated Gene Silencing in Motile T-Cells.- Live Imaging of Resident T-Cell Migration in Human Lymphoid Tissue Slices using Confocal Microscopy.- Phosphoprotein Enrichment for Protein Analysis in Motile T-Lymphocytes.- Immunometabolomic Phenotyping of Motile T-Cells.- Profiling Activity of Cellular Kinases in Migrating T-Cells.- Biotinylation Assay to Determine LFA-1 Recycling in Motile T-Lymphocytes.- A Protocol to Study T-Cell Signalling in an Immune Synapse by Microscopy.- Enzyme-Linked Immunosorbent Assay for T-Cell Dependent Immunogenicity Assessment of Therapeutic Peptides.- Modified Intravital Microscopy to Assess Vascular Health and T-Cell Motility.- Computational Analysis of Protein-Protein Interactions in Motile T-Cells.