T-Cell Motility: Methods and Protocols

Description

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 of immunology, T-cell biology, biochemistry, molecular biology, and imaging. Front Matter ....Pages i-x An Introduction to LFA-1/ICAM-1 Interactions in T-Cell Motility (Navin Kumar Verma, Dermot Kelleher)....Pages 1-9 Isolation of Human Peripheral Blood T-Lymphocytes (Atish Kizhakeyil, Seow Theng Ong, Mobashar Hussain Urf Turabe Fazil, Madhavi Latha Somaraju Chalasani, Praseetha Prasannan, Navin Kumar Verma)....Pages 11-17 A Laboratory Model to Study T-Cell Motility (Navin Kumar Verma, Shyam Mohan Palapetta, Seow Theng Ong, Mobashar Hussain Urf Turabe Fazil, Madhavi Latha Somaraju Chalasani, Praseetha Prasannan et al.)....Pages 19-23 Quantification of T-Cell Migratory Phenotypes Using High-Content Analysis (Aik Seng Ng, Seow Theng Ong, Dermot Kelleher, Navin Kumar Verma)....Pages 25-32 Live Cell Imaging and Analysis to Capture T-Cell Motility in Real-Time (Seow Theng Ong, Navin Kumar Verma)....Pages 33-40 Three-Dimensional Structured Illumination Microscopy (3D-SIM) to Dissect Signaling Cross-Talks in Motile T-Cells (Seow Theng Ong, Graham D. Wright, Navin Kumar Verma)....Pages 41-50 Real-Time Impedance-Based Detection of LFA-1-Stimulated T-Cell Transwell Chemotaxis (Praseetha Prasannan, Navin Kumar Verma)....Pages 51-57 Quantitative Real-Time PCR for Evaluating Transcriptional Changes in T-Lymphocytes (Atish Kizhakeyil, Navin Kumar Verma)....Pages 59-66 GapmeR-Mediated Gene Silencing in Motile T-Cells (Mobashar Hussain Urf Turabe Fazil, Seow Theng Ong, Madhavi Latha Somaraju Chalasani, Atish Kizhakeyil, Navin Kumar Verma)....Pages 67-73 Live Imaging of Resident T-Cell Migration in Human Lymphoid Tissue Slices Using Confocal Microscopy (Emmanuel Donnadieu, Yvonne Michel, Martin-Leo Hansmann)....Pages 75-82 Phosphoprotein Enrichment for Protein Analysis in Motile T-Lymphocytes (Praseetha Prasannan, Rathi Saravanan, Navin Kumar Verma)....Pages 83-90 Immunometabolomic Phenotyping of Motile T-Cells (Madhavi Latha Somaraju Chalasani, Gaelin Looi Wen Dong, Navin Kumar Verma)....Pages 91-98 Profiling Activity of Cellular Kinases in Migrating T-Cells (Chandra Sekhar Chirumamilla, Mobashar Hussain Urf Turabe Fazil, Claudina Perez-Novo, Savithri Rangarajan, Rik de Wijn, Padma Ramireddy et al.)....Pages 99-113 Biotinylation Assay to Determine LFA-1 Recycling in Motile T-Lymphocytes (Malin Samuelsson, Lena M. Svensson)....Pages 115-122 A Protocol to Study T-Cell Signaling in an Immune Synapse by Microscopy (Mobashar Hussain Urf Turabe Fazil, Pankaj Kumar, Navin Kumar Verma)....Pages 123-128 Enzyme-Linked Immunosorbent Assay for T-Cell Dependent Immunogenicity Assessment of Therapeutic Peptides (Madhavi Latha Somaraju Chalasani, Rajamani Lakshminarayanan, Navin Kumar Verma)....Pages 129-138 Modified Intravital Microscopy to Assess Vascular Health and T-Cell Motility (Geoffrey W. Payne, Kevin Mitchell, Stephanie L. Sellers)....Pages 139-147 Computational Analysis of Protein–Protein Interactions in Motile T-Cells (Sunil Kumar, Mobashar Hussain Urf Turabe Fazil, Khurshid Ahmad, Minaketan Tripathy, Jagath Chandana Rajapakse, Navin Kumar Verma)....Pages 149-156 Back Matter ....Pages 157-158