Main description:

This edition provides a detailed, up-to-date overview of methods used in the field of immune tolerance. Chapters guide readers through tolerogenic cell types, isolating tolerogenic cell populations for study and therapeutic utility, multiple methods to study the mechanisms underpinning tolerance, methods to induce tolerance through thymus progenitors, and methods to assess the breakdown of immune tolerance in specific pathological conditions. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Immunological Tolerance: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

PART I: Isolation of Stem Cells and Their Progeny for Immunological Tolerance Applications

1. Isolation and Characterisation of Human Adipose-derived Stem Cells

Wilson, A., Chee, M.J., Butler P., and Boyd, A.S.

2. Isolation of Murine Hematopoietic Stem Cells

Juan Menendez-Gonzalez, Lubaid Saleh, Gui Jie Feng and Neil P. Rodrigues

3. Generation of Antigen-specific T cells from Human Induced Pluripotent Stem Cells Toshinobu Nishimura and Hiromitsu Nakauchi

PART II: Isolation of Regulatory Lymphocyte Populations

4. Isolation of Human Regulatory T Lymphocytes by Fluorescence-activated Cell Sorting

Kate Milward, Joanna Hester, and Kathryn J Wood

5. Identification and isolation of Regulatory B cells in Mouse and Human

Madhvi Menon, Elizabeth C. Rosser, and Claudia Mauri

6. An innovative Standard Operation Procedure for Isolating GMP-grade CD4+CD25+ T Cells from Non-mobilized Leukapheresis

W. Zhang, S. M. Watt, and D. J. Roberts

PART III: Mechanisms and Induction of Immunological Tolerance

7. Measuring CTLA-4-dependent Suppressive Function in Regulatory T Cells

Tie Zheng Hou, Omar S Qureshi, and David M. Sansom

8. Alloanergization Method for Inducing Allospecific Hyporesponsiveness in PBMC Exposed to Allostimulation in vitro in the Context of Costimulatory Molecule Blockade

Christine M. Barbon, Kenneth J. Janec Rowan H. Kelner, James E. Norton, and Eva C. Guinan

9. In Vitro Suppression of CD4+ T Cell Responses by Murine and Human Myeloid-derived Suppressor Cells

Katerina Vlachou and Panayotis Verginis

10. Disassembling and Re-aggregating the Thymus: The Plus and Minus of Current Assays

Elia Piccinini and Paola Bonfanti

11. Isolation of Highly Viable Thymic Epithelial Cells for use in In Vitro and In Vivo Experiments

Mi-Jeong Kim and Thomas Serwold

PART IV: Transplantation Tolerance

12. Immune Tolerance and Rejection in Organ Transplantation

Jessica Stolp, Masaaki Zaitsu, and Kathryn J Wood

13. Method of Generating Tolerogenic Maturation-resistant Dendritic Cells and Testing for their Immune-regulatory Functions In Vivo in the Context of Transplantation

Sherrie J. Divito and Adrian E. Morelli

PART V: The Breakdown of Immunological Tolerance in Disease

14. Luminex Coupled-EliFACS: A Multiparametric Method to Enumerate and Functionally Characterize Autoreactive T Cells in Human Peripheral Blood

HuiJuan Lv, Khadir Raddassi, and Myra A Lipes

15. Clinical and Functional Evaluation of Ocular Inflammatory Disease Using the Model of Experimental Autoimmune Uveitis Jun Chen, and Rachel R. Caspi

16. Methods of Isolation and Analysis of TREG Immune Infiltrates from Injured and Dystrophic Skeletal Muscle

Burzyn Dalia, Wagers Amy, Mathis Diane, and Cerletti Massimiliano