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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.

T-Cell Receptor Signaling: Methods and Protocols

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