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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.
List of contents
Human and Murine T helper Cell Recovery from Organs and Tissues.- Identification and Purification of Human Memory T Helper Cells from Peripheral Blood.- Transcription Factor Evaluation by Flow Cytometry.- Mass Cytometry Analysis of T Helper Cells.- Murine T Helper Cell Differentiation and Plasticity.- Identification of Follicular T Cell Subsets in Murine and Human Tissues.- Analysis of T Cells In Inflamed Non-Lymphoid Tissues.- TCR Vbeta Evaluation by Flow Cytometry.- Evaluation of Cytokine Production at the Single Cell Level by Flow Cytometry Upon Polyclonal Stimulation.- Evaluation of Secreted Cytokines by Multiplex Bead Based Assay (X MAP Technology, Luminex).- Quantifying Antigen-specific T Cells by Assessing their Antigen-Induced Proliferation.- Flow Cytometric Characterization of Human Antigen-Reactive T helper Cells.- Identification of Human Antigen-Specific CD4+ cells with Peptide-MHC Multimer Technologies.- Human Tcell Cloning by Limiting Dilution.- Human Germinal Center Reaction: Assessing T Follicular Helper-B cell Interactions In Vitro.- Study of Signal Transduction Pathways by Phospho-Protein Evaluation.- The Study of Protein-DNA Interactions in CD4+ T Cells using ChIPmentation.- mRNA Transfection of T lymphocytes by Electroporation.- Transduction of Human T Cell Subsets with Lentivirus.- Plasmid- or Ribonucleoprotein-mediated CRISPR/Cas Gene Editing in Primary Murine T Cells.- Generation of Sequencing Libraries for Building Immune Cell Methylomes.- Gene Expression Analysis of T Cells by Single Cell RNA-seq.- Quantifying Proteome and Protein Modifications in Activated T Cells by Multiplexed Isobaric Labeling Mass Spectrometry.- Metabolomics, Lipidomics and Immunometabolism.- Experimental Mouse Models of Asthma and Analysis of CD4+ T Cells.- Murine T Cell Transfer Colitis as a Model for Inflammatory Bowel Disease.- ExperimentalAutoimmune Encephalomyelitis.