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. 

Part 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

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

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

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