Kearse T Cell Protocols

Development and Activation
Erscheinungsjahr 2008
ISBN: 978-1-59259-682-9
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 365 Seiten

Reihe: Springer Protocols

The purpose of T Cell Protocols: Development and Activation is to c- lect a series of protocols, particularly those that have been developed within the past few years, to help investigators master new techniques (or improve existing ones) for the study of T-cell Biology. Invariably, in putting together a book like this it is difficult to decide which methods to include and which to leave out. To this end methods were selected from a variety of disciplines, including cellular immunology, b- chemistry, and molecular biology, to try to provide something of interest for everyone who works on T-cell development and activation. I would like to mention that my primary reason for agreeing to put this book together is that, when I was a graduate student, I purchased a copy of Selected Methods in Cellular Immunology by Mishell and Shigii which proved a tremendous help in learning the basics of one-and two- dimensional gel te- niques (and other methods). The cover has long since fallen off, but it still remains one of my most valued reference books for the laboratory. It is my hope that T Cell Protocols: Development and Activation will prove similarly useful to current and future scientists wishing to learn new methods for expl- ing the development and activation of T cells.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Kearse, Kelly P.

Inhaltsverzeichnis

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 TyrosinePhosphatase 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).