T-Cell Motility, 1st ed. 2019 Methods and Protocols Methods in Molecular Biology Series, Vol. 1930

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.

Preface…
Table of Contents…
Contributing Authors…

1. An Introduction to LFA-1/ICAM-1 Interactions in T-Cell Motility
Navin Kumar Verma and Dermot Kelleher

2. Isolation of Human Peripheral Blood T Lymphocytes
Atish Kizhakeyil, Seow Theng Ong, Mobashar Hussain Urf Turabe Fazil, Madhavi Latha Somaraju Chalasani, Praseetha Prasannan, and Navin Kumar Verma

3. A Laboratory Model to Study T-Cell Motility
Navin Kumar Verma, Sjyam Mohan Palapetta, Seow Theng Ong, Mobashar Hussain Urf Turabe Fazil, Madhavi Latha Somaraju Chalasani, Praseetha Prasannan, Atish Kizhakeyil, and Dermot Kelleher

4. Quantification of T-Cell Migratory Phenotypes using High Content Analysis
Aik Seng Ng, Seow Theng Ong, Dermot Kelleher, and Navin Kumar Verma

5. Live Cell Imaging and Analysis to Capture T-Cell Motility in Real-Time
Seow Theng Ong and Navin Kumar Verma

6. Three-Dimensional Structured Illumination Microscopy (3D-SIM) to Dissect Signalling Cross-Talks in Motile T-Cells
Seow Theng Ong, Graham D. Wright, and Navin Kumar Verma

7. Real-Time Impedance-Based Detection of LFA-1-Stimulated T-Cell Transwell Chemotaxis
Praseetha Prasannan and Navin Kumar Verma

8. Quantitative Real-Time PCR for Evaluating Transcriptional Changes in T-Lymphocytes
Atish Kizhakeyil and Navin Kumar Verma

9. GapmeR-Mediated Gene Silencing in Motile T-Cells
Mobashar Hussain Urf Turabe Fazil, Seow Theng Ong, Madhavi Latha Chalasani, Atish Kizhakeyil, and Navin Kumar Verma

10. Live Imaging of Resident T-Cell Migration in Human Lymphoid Tissue Slices using Confocal Microscopy
Emmanuel Donnadieu, Yvonne Michel, and Martin-Leo Hansmann

11. Phosphoprotein Enrichment for Protein Analysis in Motile T-Lymphocytes
Praseetha Prasannan, Rathi Saravanan, and Navin Kumar Verma

12. Immunometabolomic Phenotyping of Motile T-Cells
Madhavi Latha Somaraju Chalasani, Gaelin Looi Wen Dong, and Navin Kumar Verma

13. 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, Navin Kumar Verma, and Wim Vanden Berghe

14. Biotinylation Assay to Determine LFA-1 Recycling in Motile T-Lymphocytes
Malin Samuelsson and Lena M. Svensson

15. A Protocol to Study T-Cell Signalling in an Immune Synapse by Microscopy
Mobashar Hussain Urf Turabe Fazil, Pankal Kumar, and Navin Kumar Verma

16. Enzyme-Linked Immunosorbent Assay for T-Cell Dependent Immunogenicity Assessment of Therapeutic Peptides
Madhavi Latha Somaraju Chalasani, Rajamani Lakshminarayanan, and Navin Kumar Verma

17. Modified Intravital Microscopy to Assess Vascular Health and T-Cell Motility
Geoffrey W. Payne, Kevin Mitchell, and Stephanie L. Sellers

18. Computational Analysis of Protein-Protein Interactions in Motile T-Cells
Sunil Kumar, Mobashar Hussain Urf Turabe Fazil, Khursheed Ahmad, Minaketan Tripathy, Jagath Chandana Rajapakse, and Navin Kumar Verma

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts