Main description:

The aim of this volume is to provide a comprehensive description of methods and protocols useful for the further study of T-helper cells. Chapters guide readers through T-helper cell recovery, molecular study, signal transduction pathways, T-cell manipulation and, last but not least, "omic" approaches. 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- Helper Cells: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.

Contents:

1. Human and Murine T-Helper Cell Recovery from Organs and Tissues

Francesco Siracusa, Annette Lahmann, Anna Vanni, Filippo Cortesi, and Alessio Mazzoni

2. Identification and Purification of Human Memory T-Helper Cells from Peripheral Blood

Alessia Capone and Elisabetta Volpe

3. Transcription Factor Evaluation by Flow Cytometry

Daniela Fenoglio and Tiziana Altosole

4. Mass Cytometry Analysis of T-Helper Cells

Priyanka B. Subrahmanyam and Holden T. Maecker

5. Murine T-Helper Cell Differentiation and Plasticity

Francesco Siracusa, Franziska Muscate, and Laura Garcia Perez

6. Identification of Follicular T-Cell Subsets in Murine and Human Tissues

Annette Lahmann, Laura Bauer, and Andreas Hutloff

7. Analysis of T-Cells In Inflamed Non-Lymphoid Tissues

Laura Bauer and Andreas Hutloff

8. TCR V Evaluation by Flow Cytometry

Benedetta Peruzzi, Sara Bencini, and Roberto Caporale

9. Evaluation of Cytokine Production at the Single Cell Level by Flow Cytometry Upon Polyclonal Stimulation

Jakob Zimmermann

10. Evaluation of Secreted Cytokines by Multiplex Bead Based Assay (X MAP Technology, Luminex)

Letizia Lombardelli, Federica Logiodice, Ornela Kullolli, and Marie-Pierre Piccinni

11. Quantifying Antigen-specific T-Cells by Assessing their Antigen-Induced Proliferation

Aayushi Shah and Hyun-Dong Chang

12. Flow Cytometric Characterization of Human Antigen-Reactive T-Helper Cells

Carina Saggau, Alexander Scheffold, and Petra Bacher

13. Identification of Human Antigen-Specific CD4+ cells with Peptide-MHC Multimer Technologies

I-Ting Chow and William W. Kwok

14. Human T-cell Cloning by Limiting Dilution

Laura Maggi, Manuela Capone, and Alessio Mazzoni

15. Human Germinal Center Reaction: Assessing T-Follicular Helper-B cell Interactions In Vitro

Emeline Thevenon, Vanessa Cornacchione, Elisabetta Traggiai, and Francesca Rucci

16. Study of Signal Transduction Pathways by Phospho-Protein Evaluation Manuela Capone, Alessio Mazzoni, and Laura Maggi

17. The Study of Protein-DNA Interactions in CD4+ T Cells using ChIPmentation

Arnulf Hertweck and Richard G. Jenner

18. mRNA Transfection of T lymphocytes by Electroporation

Lea-Isabell Schwarze and Boris Fehse

19. Transduction of Human T-Cell Subsets with Lentivirus

Vivian C. W. Fung, Isaac Rosado-Sanchez, Megan K Levings

20. Plasmid- or Ribonucleoprotein-mediated CRISPR/Cas Gene Editing in Primary Murine T-Cells

Marianne Doelz, Romina Marone, and Lukas T. Jeker

21. Generation of Sequencing Libraries for Building Immune Cell Methylomes

Stefan Floess and Jochen Huehn

22. Gene Expression Analysis of T-Cells by Single Cell RNA-seq

Domenico Lo Tartaro, Sara De Biasi, Mattia Forcato, Lara Gibellini, and Andrea Cossarizza

23. Quantifying Proteome and Protein Modifications in Activated T-Cells by Multiplexed Isobaric Labeling Mass Spectrometry

Haiyan Tan, Daniel Bastardo Blanco, Boer Xie, Yuxin Li, Zhiping Wu, Hongbo Chi, and Junmin Peng

24. Metabolomics, Lipidomics and Immunometabolism

Fortunata Carbone, Sara Bruzzaniti, Clorinda Fusco, Alessandra Colamatteo, Teresa Micillo, Paola de Candia, Fabrizia Bonacina, Giuseppe Danilo Norata and Giuseppe Matarese

25. Experimental Mouse Models of Asthma and Analysis of CD4+ T Cells

William J. Branchett, Simone A. Walker, and Clare M. Lloyd

26. Murine T-Cell Transfer Colitis as a Model for Inflammatory Bowel Disease

Patrick Maschmeyer, Jakob Zimmermann, and Anja Andrea Kuhl

27. Experimental Autoimmune Encephalomyelitis

Clara Ballerini