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Driven by methodological success in identifying reliable lineage markers, regulatory T cells have quickly been recognized as the most numerous subset of immune regulators in the body with critical functions in a wide array of immune responses. In Regulatory T Cells: Methods and Protocols, experts in the field to offer a collection of current techniques to advance the study of regulatory T cells, including the use of the IL-2 receptor alpha chain and other markers, as well as the more recently desirable use of the transcription factor FoxP3. Divided into three sections, the book covers, in equal measure, in vitro, in vivo, and human studies. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known methodological pitfalls. Authoritative and cutting-edge, Regulatory T Cells: Methods and Protocols distills the most vital current techniques through several years of optimization and standardization in order to allow reliable and reproducible use by both young and experienced cellular and molecular immunologists.

Written in the popular Methods in Molecular Biology™ series format, this book covers, in equal measure, in vitro, in vivo, and human studies in regulatory T cells, offering readily reproducible laboratory protocols, key tips on troubleshooting and more.

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Part I: Introduction

1. Regulatory T Cells: History and Perspective
Shimon Sakaguchi

Part II: In vitro

2. In vitro Treg Suppression Assays
Lauren W. Collison and Dario A.A. Vignali

3. Generation of T Cell Hybridomas from Naturally-Occurring FoxP3+ Regulatory T Cells Nagendra Singh, Rafal Pacholczyk, Makio Iwashima, and Leszek Ignatowicz

4. In vitro and In vivo Analyses of Regulatory T Cell Suppression of CD8+ T Cells
Kim J. Hasenkrug and Lara M. Myers

5. Flow Cytometric Profiling of Mature and Developing Regulatory T Cells in the Thymus
Donald M. Simons and Andrew J. Caton

6. ChIP-on-Chip for FoxP3
Ye Zheng

7. Live Imaging of Dendritic Cell-Treg Cell Interactions
Milka Sarris and Alexander G. Betz

Part III: In vivo

8. Genetic Tools for Analysis of FoxP3+ Regulatory T Cells In vivo
Nadia M. Jeremiah and Adrian Liston

9. In vivo Treg Suppression Assays
Creg J. Workman, Lauren W. Collison, Maria Bettini, Meenu R. Pillai, Jerold E. Rehg, and Dario A.A. Vignali

10. In vivo Depletion of FoxP3+ Tregs Using the DEREG Mouse Model
Katharina Lahl and Tim Sparwasser

11. Antigen-Specific Induction of Regulatory T Cells In vivo and In vitro
Carolin Daniel, Hidde Ploegh, and Harald von Boehmer

12. In vitro Expansion of Alloantigen-Specific Regulatory T Cells and Their Use in Prevention of Allograft Rejection
Clemence Nouze, Lise Pasquet, and Joost P.M. van Meerwijk

Part IV: Human

13. Analysis of Human FoxP3+ Treg Cells Phenotype and Function
Eva d'Hennezel and Ciriaco A. Piccirillo

14. Depletion of Human Regulatory T Cells
Amy C. Hobeika, Michael A. Morse, Takuya Osada, Sharon Peplinski, H. Kim Lyerly, and Timothy M. Clay

15. Assessment of Suppressive Capacity by Human Regulatory T Cells Using a Reproducible, Bi-Directional CFSE-Based In vitro Assay
Anya Schneider and Jane H. Buckner

16. Measurement of Proliferation and Disappearance of Regulatory T Cells in Human Studies Using Deuterium-Labeled Glucose
Milica Vukmanovic-Stejic, Yan Zhang, Arne N. Akbar, and Derek C. Macallan

17. Flow Cytometric Detection of Human Regulatory T Cells
Barbara Fazekas de St. Groth, Erhua Zhu, Suzanne Asad, and Loretta Lee

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