Regulatory T-Cells Methods and Protocols

This volume provides methods and models for investigating the immunosuppressive subset of CD4+ T-cells, regulatory T-cells (Treg). Chapters guide readers through, basic protocols for analyzing Treg in mice and humans, single-cell analysis for analyzing Treg, and methods for the analysis of Treg in clinical application. 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, Regulatory T-Cells: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. 

Analysis of the in vivo function of follicular regulatory T (TFR) cells in the regulation of antibody response.- Analysis of peripherally derived Treg in the intestines.- Isolation and analysis of tumor infiltrating Treg.- Induction of peripheral Treg.- Adoptive transfer and bone marrow chimera models to analyse Treg function and differentiation.- Functional analysis of Foxp3 and its mutants by retroviral transduction of murine primary CD4⁺ T cells.- Genetic Tools for Analyzing Foxp3⁺ Treg cells: Fluorochrome-Based Transcriptional Reporters and Genetic Fate-Mapping.- Depletion of Treg by the Diphtheria Toxin System.- Analysis of FOXP3 DNA methylation patterns to identify functional FOXP3+ T-Cell subpopulations.- Protein Ubiquitination Assay.- Phenotypic and Functional Studies of Human Treg cell Subpopulations.- In vitro analysis of CTLA-4 mediated transendocytosis by Regulatory T cells.- Isolation and functional characterisation of regulatory CD4+ Tcells from the inflamed joints of patients with rheumatoid arthritis.- A GMP protocol for the manufacture of regulatory T cells for clinical application.- Regulatory T-cell phenotyping using CyTOF.- High-dimensional single-cell profiling of tumor-infiltrating CD4+ regulatory T cells.- Single-cell transcriptome analysis of Treg.