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This volume provides current and new advanced methods and protocols to study T cells. Chapters guide readers through T cell diversity using mass cytometry, analyzing T cells from single cell level, CRISPR/Cas9 techniques to study the T cell activation, techniques to study subsets of Tcell’s, procedures to study artificial antigen presentosomes for T cell activation, techniques to study the T cell development, two-photon microscopy, and MAIT cells. 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, T-Cell Receptor Signaling: Methods and Protocols aims to provide a wide range of approaches and be an invaluable resource for present and future generations of T cell researchers.
1. Exploration of T Cell Diversity using Mass Cytometry
Kaitlin C O’Boyle, Takuya Ohtani, Bertram Bengsch, Sasikanth Manne, Sarah E Henrickson, E John Wherry, and Cecile Alanio
2. Analysis of Transcriptional Profiling of Immune Cells at the Single-cell Level
Annabel Ferguson and Kong Chen
3. Simultaneous Measurement of Surface Proteins and Gene Expression from Single Cells
Jiadi Luo, Carla A. Erb, and Kong Chen
4. A Carrier Strategy for Mass Cytometry Analysis of Small Numbers of Cells
Xian Jia, Xiaojuan Zhou, Haiping Zheng, Shan Jiang, Jiannan Weng, Lei Huang, Zhiqiang Du, Changchun Xiao, Lei Zhang, Xiao Lei Chen, and Guo Fu
5. CRISPR/Cas9-based Genetic Screening to Study T Cell Function
Wanjing Shang, Fei Wang, Qi Zhu , Liangyu Wang, and Haopeng Wang
6. Preferential Expansion of CD4+Foxp3+ Regulatory T Cells (Tregs) In Vitro by Tumor Necrosis Factor
Chon-Kit Chou and Xin Chen
7. CD4+ T Cell Differentiation in vitro
Wenyong Yang, Xueying Chen, and Hongbo Hu
8. Characterization of Immune Cell Subset Expansion in Response to Therapeutic Treatment in Mice
Jakub Tomala and Jamie B. Spangler
9 Primary T Cell Transduction to Study Follicular Helper T Cell Differentiation
Yang Zhang, Xuehui Long, and Xiaoming Wang
10. In vitro Generation of Stem Cell Memory-like T Cells from Activated T Cells
Makoto Ando, Mari Ikeda and Akihiko Yoshimura, and Taisuke Kondo
11. Artificial Antigen Presentosomes for T Cell Activation
Yi-Geng Pang, Chien-Chung Chang
12. Imaging Chimeric Antigen Receptor (CAR) -triggered T Cell Activation
Kendra Libby and Xiaolei Su
13. Assessing the Impact of Phytochemicals on Immune Checkpoints: Implications for Cancer Immunotherapy
Melanie R. Power Coombs and David W. Hoskin
14. Testing the Efficiency and Kinetics of Negative Selection Using Thymic Slices
Tyng-An Zhou, Chia-Lin Hsu, and Ivan Lilyanov Dzhagalov
15. Investigating T Cell Receptor Signaling Dynamics in situ by Two-photon Microscopy of Thymocytes Expressing Genetic Reporters in Low-Density Chimeras
Marilaine Fournier, Mengqi Dong, and Heather J. Melichar
16. Retroviral Gene Transduction into T-cell Progenitors for Analysis of T Cell Development in the Thymus
Ryunosuke Muro, Hiroshi Takayanagi, and Takeshi Nitta
17. An Integrated Strategy for Identifying Targets of Ubiquitin-mediated Degradation in CD4+ T Cells
Natania S. Field, Claire E. O’Leary, Joseph M. Dybas, Hua Ding, and Paula M. Oliver
18. Radioisotope Based Protocol for Determination of Central Carbon Metabolism in T Cells
Xuyong Chen, John Sherman, and Ruoning Wang
19. Assessment of Immune Protective T cell Repertoire in Humans Immunized with Novel Tuberculosis Vaccines
Mangalakumari Jeyananthan and Zhou Xing
20. Studying Peripheral T Cell Homeostasis in Mice; A Concise Technical Review
Moutuaata M. Moutuou, Simon-David Gauthier, Nicolas Chen, Dominique Leboeuf, and Martin Guimond
21. Detection, Expansion, and Isolation of Human MAIT Cells
Yu Liu, Wei Wang, Xiongwen Wu and Xiufang Weng
22. In vitro Differentiation of CD4+ T Cell Effector and Regulatory Subsets
Jaclyn R. Espinosa, Joshua D. Wheaton, and Maria Ciofani
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