Overview: For Chapters 2 - 15 -- Insights into T-Cell Development Using Transgenic and Knockout Mice -- T Cell Development -- Isolation and Characterization of Murine Early Intrathymic Precursor Populations -- Differentiation of Mouse Thymocytes in Fetal Thymus Organ Culture -- Purification of Immature CD4+CD8+ Thymocytes by Panning with Anti-CD8 Antibody -- Transfection and Transcription of Genes in Developing Thymocytes -- Analysis of CD4/CD8 Lineage Commitment by Pronase Treatment and Reexpression Assay -- Intrathymic Delivery of MHC Genes Using Recombinant Adenoviruses -- Quantitative Analysis of the Usage of Human T-Cell Receptor ? and ? Chain Variable Regions by Reverse Dot Blot Hybridization -- Flow Cytometric Detection of Cytoplasmic and Surface CD3-? Expression in Developing T Cells -- Assessing Apoptosis of Developing T Cells by Flow Cytometry -- Isolation of T-Cell Subsets by Magnetic Cell Sorting (MACS) -- Isolation of T-Cell Antigens by Retrovirus-Mediated Expression Cloning -- Flow Cytometric Analysis of Murine T Lymphocytes -- Production and Characterization of Immature Murine T-Lymphoma Cell Lines -- Production and Characterization of T-Cell Hybridomas -- Overview for Chapters 17-29 -- Something Happens; A Brief History of TCR Signal Transduction -- T-Cell Activation -- Measurement of Phosphatidylinositol (PI) Hydrolysis in Activated T Lymphocytes -- Fluorescence Polarization as an Early Measure of T-Lymphocyte Stimulation -- Measurement of Lymphoproliferation at the Single-Cell Level by Flow Cytometry -- Digital Image Analysis of Lymphocyte Activation -- Induction of T-Cell Signaling by Immobilized Integrin Ligands -- Cytotoxic T-Cell Adherence Assay (CAA) -- Detecting Ubiquitinated T-Cell Antigen Receptor Subunits by Immunoblotting -- Determination of CD45 Tyrosine Phosphatase Activity in T Lymphocytes -- Measurement of Protein Tyrosine Phosphorylation in T-Cell Subsets by Flow Cytometry -- Biochemical Analysis of Activated T Lymphocytes -- Activation of Heterotrimeric GTP-Binding Proteins Upon TCR/CD3 Engagement -- Generation of Human T Helper 1 and T Helper 2 Subsets from Peripheral Blood-Derived Naive CD4+ T Cells -- Transcriptional Expression Analysis of T-Cell Activation by Multiplex Messenger Assay (MMA).

In T Cell Protocols: Development and Activation, Kelly P. Kearse and a panel of laboratory experts describe the step-by-step execution of a wide variety of basic and specialized techniques for the study of T cell developmental biology and of their subsequent activation. Readily reproducible, the techniques are explained concisely, but in such detail that they are suitable for both the novice and experienced investigator. Coverage includes treatment of both fundamental and novel experimental protocols for the analysis of the role T cells play in immune function and in the prevention and cure of human diseases. Practical and timely, T Cell Protocols: Development and Activation offers basic scientists and clinical investigators a valuable compendium of key protocols that will improve skills with existing methods, as well as afford rapid mastery of powerful new techniques.