Description
Book SynopsisThis 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.
Table of ContentsPart I Immunological techniques for analysing Treg
1. Analysis of the in vivo function of follicular regulatory T (TFR) cells in the regulation of antibody response
Lin Luo and Jianmei W. Leavenworth
2. Analysis of peripherally derived Treg in the intestines
Tadashi Takeuchi and Hiroshi Ohno
3. Isolation and analysis of tumor infiltrating Treg
Katharina Bergerhoff
4. Induction of peripheral Treg
José Almeida-Santos, Marie-Louise Bergman, and Jocelyne Demengeot
5. Adoptive transfer and bone marrow chimera models to analyse Treg function and differentiation
Vital da Silva Domingues, Iris Caramalho, Marie-Louise Bergman, and Jocelyne Demengeot
Part III Molecular techniques for analysing Treg
6. Functional analysis of Foxp3 and its mutants by retroviral transduction of murine primary CD4+ T cells
Akira Nakajima, Ryuichi Murakami, Shohei Hori
7. Genetic Tools for Analyzing Foxp3+ Treg cells: Fluorochrome-Based Transcriptional Reporters and Genetic Fate-Mapping
Mario Simonetti, Acelya Yilmazer, and Karsten Kretschmer
8. Depletion of Treg by the Diphtheria Toxin System
Sho Hanakawa, Zachary Chow, and Kenji Kabashima
9. Analysis of FOXP3 DNA methylation patterns to identify functional FOXP3+ T-Cell subpopulations
Ekaterina Minskaia and João F. Lacerda
10. Protein Ubiquitination Assay
Ping Wei, Rui Liang, and Fan Pan
Part IV Human Treg analysis and clinical application
11. Phenotypic and Functional Studies of Human Treg cell Subpopulations
Mo Atif, Mustapha Cherai, Makoto Miyara
12. In vitro analysis of CTLA-4 mediated transendocytosis by Regulatory T cells
Erin Waters, Cayman Williams, Alan Kennedy, and David M Sansom
13. Isolation and functional characterisation of regulatory CD4+ T cells from the inflamed joints of patients with rheumatoid arthritis
Giovanni A.M. Povoleri, Veerle Fleskens and Leonie S. Taams
14. A GMP protocol for the manufacture of regulatory T cells for clinical application
Alice O’Neil, Matthew Brook, Seetha Abdul-Wahab, Joanna Hester, Giovanna Lombardi and Fadi Issa
Part V Single cell and high-dimensional techniques for analysing Treg
15. Regulatory T-cell phenotyping using CyTOF
Hugo Barcenilla, Mikael Pihl, Florence Sjögren, Louise Magnusson, and Rosaura Casas
16. High-dimensional single-cell profiling of tumor-infiltrating CD4+ regulatory T cells
Giorgia Alvisi, Simone Puccio, Rahul Roychoudhuri, Caterina Scirgolea and Enrico Lugli
17. Single-cell transcriptome analysis of Treg
Benjy Tan Jek Yang, Masahiro Ono, and Yorifumi Satou
