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. 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
