T Cell Activation Methods and Protocols

This volume provides detailed, up-to-date methods used in basic and applied research on T-cell activation. Chapters explore the fundamentals in T-cell biology a board range cutting-edge techniques from initial Ca2+ live-cell imaging to downstream effector functions in autoimmune disease. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, T- cell activation: Methods and Protocols  will be an indispensable protocol collection for researchers studying T-cell activation in inflammation and immunity.

Imaging protocol for super-resolution microscopy in fixed T cells.- Visualization and quantification of NFAT1 translocation in T cells.- Deep Learning-Based Image Restoration and Super-Resolution for Fluorescence Microscopy: Overview and Resources.- Analysing Point Accumulation in Nanoscale Topography (PAINT) data to decipher T cell receptor cluster signalling.- Measuring NAADP-evoked Ca2+ release in permeabilized T cells.- An improved two-dimensional HPLC method for endogenous 2'-deoxy-ADPR and 2'-deoxy-NAD.- Analysis of Store-Operated Ca2+ Entry in primary T cells.- Monitoring Dynamic of cAMP Signaling in Primary Murine CD4+ T Cells via Live-Cell Imaging.- Using specific reporter mice to assess CD4+ T cell transdifferentiation.- Generation of antigen-specific CD4+ primary rat T cell lines.- Induction and assessment of activation in rat effector CD4+ memory Th cells.- Imaging BiTE-mediated activation of primary human CD8+ T cells.- Flow cytometric analysis of mouse brain CD8+ T cell activation.- Utilizing pMHC Multimers to Study Antigen-Specific T Cells and T Cell Exhaustion in Chronic Viral Infections.- Analysis of TF/DNA binding in T cells applying Alpha technology.- Using Seahorse technology as an efficient way of verifying T-cell stimulation.- Metabolic screening of T lymphocytes during activation via SEAHORSE extracellular flux (XF) analysis.-  Metabolic profiling of activated T lymphocytes using single-cell energetic metabolism by profiling translation inhibition (SCENITH).