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T-Cell Differentiation

Methods and Protocols

  • Book
  • © 2017

Overview

Editors:
  1. Enrico Lugli

    1. Laboratory of Translational Immunology, Humanitas Clinical & Research Center, Rozzano, Italy

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  • Includes cutting-edge methods and protocols
  • Provides step-bystep detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1514)

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About this book

This volume provides protocols to successfully apply cutting-edge technologies to characterize the biology of T cells at an unprecedented level of complexity. Chapters guide readers through flow cytometry and fluorescence-activated cell sorting, the behaviour of single T cells after adoptive cell transfer (ACT), single cell gene expression by multiplex PCR, lentiviral transduction approaches, protocols to derive large numbers of early-differentiated memory T cells by using dedicated cytokines cocktails, approaches to measure telomerase activity in terminally differentiated T cells, and approaches to define Treg cells at the phenotypic and functional level. The final part of the book is dedicated to the analysis of the differentiation and effector functions of innate T cells, namely the well-known γ/δ T cells, and the recently identified CD8+ mucosal associated invariant T (MAIT) cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to theirrespective 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 Differentiation: Methods and Protocols aims to provide protocols that are fundamental to monitor the T cell compartment at the level of single cells in pathological and immunotherapy conditions.

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Table of contents (18 protocols)

  1. Front Matter

    Pages i-xi
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  2. The Vast Universe of T Cell Diversity: Subsets of Memory Cells and Their Differentiation

    • Camilla Jandus, Amaia Martínez Usatorre, Selena Viganò, Lianjun Zhang, Pedro Romero
    Pages 1-17

  3. Basic Aspects of T Helper Cell Differentiation

    • Nicola Gagliani, Samuel Huber
    Pages 19-30

  4. FACS Analysis of Memory T Lymphocytes

    • Enrico Lugli, Veronica Zanon, Domenico Mavilio, Alessandra Roberto
    Pages 31-47

  5. Intravital Microscopy Analysis of Hepatic T Cell Dynamics

    • Alexandre Pierre Benechet, Lucia Ganzer, Matteo Iannacone
    Pages 49-61

  6. Analysis of T Cell Activation by Confocal Microscopy

    • Paola Larghi, Antonella Viola, Barbara Molon
    Pages 63-81

  7. Phenotypic and Functional Analysis of Antigen-Specific T Cell Exhaustion

    • Kristin L. Boswell, Takuya Yamamoto
    Pages 83-92

  8. pMHC Multiplexing Strategy to Detect High Numbers of T Cell Responses in Parallel

    • Daisy Philips, Marlous van den Braber, Ton N. Schumacher, Pia Kvistborg
    Pages 93-101

  9. Differentiation of Diverse Progenies of Memory T Cells from Naïve CD8+ T Cell Precursors

    • Veronica Zanon, Enrico Lugli
    Pages 103-110

  10. Gammaretroviral Production and T Cell Transduction to Genetically Retarget Primary T Cells Against Cancer

    • Gongbo Li, Kyungho Park, Marco L. Davila
    Pages 111-118

  11. Measuring Telomerase Activity in Senescent Human T Cells Upon Genetic Modification

    • Alessio Lanna
    Pages 119-126

  12. Strategies for T Helper Cell Subset Differentiation from Naïve Precursors

    • Francesco Annunziato, Laura Maggi
    Pages 127-137

  13. Phenotypic and Functional Analysis of the Suppressive Function of Human Regulatory T Cells

    • Eleonora Timperi, Vincenzo Barnaba, Silvia Piconese
    Pages 139-151

  14. Approaches to Detect microRNA Expression in T Cell Subsets and T Cell Differentiation

    • Silvia Monticelli, Tarmo Äijö, Sara Trifari
    Pages 153-172

  15. Next-Generation Sequencing Analysis of Long Noncoding RNAs in CD4+ T Cell Differentiation

    • Valeria Ranzani, Alberto Arrigoni, Grazisa Rossetti, Ilaria Panzeri, Sergio Abrignani, Raoul J. P. Bonnal et al.
    Pages 173-185

  16. Highly Multiplexed, Single Cell Transcriptomic Analysis of T-Cells by Microfluidic PCR

    • Maria Dominguez, Mario Roederer, Pratip K. Chattopadhyay
    Pages 187-202

  17. Single-Cell RNA Sequencing of Human T Cells

    • Alexandra-Chloé Villani, Karthik Shekhar
    Pages 203-239

  18. Extensive Phenotypic Analysis, Transcription Factor Profiling, and Effector Cytokine Production of Human MAIT Cells by Flow Cytometry

    • Joana Dias, Johan K. Sandberg, Edwin Leeansyah
    Pages 241-256

  19. Developmental and Functional Assays to Study Murine and Human γδ T Cells

    • Julie C. Ribot, Karine Serre, Bruno Silva-Santos
    Pages 257-267

  20. Back Matter

    Pages 269-271
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Editors and Affiliations

  • Laboratory of Translational Immunology, Humanitas Clinical & Research Center, Rozzano, Italy

    Enrico Lugli

Bibliographic Information

  • Book Title: T-Cell Differentiation

  • Book Subtitle: Methods and Protocols

  • Editors: Enrico Lugli

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-6548-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2017

  • Hardcover ISBN: 978-1-4939-6546-5Published: 28 October 2016

  • Softcover ISBN: 978-1-4939-8233-2Published: 27 June 2018

  • eBook ISBN: 978-1-4939-6548-9Published: 27 October 2016

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 271

  • Number of Illustrations: 11 b/w illustrations, 44 illustrations in colour

  • Topics: Immunology

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