About the Book
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.
Table of Contents:
Part I. Insights into T-Cell Development Using Transgenic and Knockout Mice, M. Albert Basson and Rose Zamoyska. Part II. T Cell Development Isolation and Characterization of Murine Early Intrathymic Precursor Populations, Li Wu and Ken Shortman. Differentiation of Mouse Thymocytes in Fetal Thymus Organ Culture,Yousuke Takahama. Purification of Immature CD4+CD8+ Thymocytes by Panning with Anti-CD8 Antibody, Terrence G. Gardner and Kelly P. Kearse. Transfection and Transcription of Genes in Developing Thymocytes, Alison M. Michie and Juan Carlos Zuniga-Pflucker. Analysis of CD4/CD8 Lineage Commitment by Pronase Treatment and Reexpression Assay, Harumi Suzuki. Intrathymic Delivery of MHC Genes Using Recombinant Adenoviruses, Ronald Rooke, Christophe Benoist, and Diane Mathis. Quantitative Analysis of the Usage of Human T-Cell Receptor a and b Chain Variable Regions by Reverse Dot Blot Hybridization, Takaji Matsutani, Takeshi Yoshioka, Yuji Tsuruta, Shoji Iwagami, Tomoko Toyosaki-Maeda, and Ryuji Suzuki. Flow Cytometric Detection of Cytoplasmic and Surface CD3-e Expression in Developing T Cells, Karl J. Franek and Robert Chervenak. Assessing Apoptosis of Developing T Cells by Flow Cytometry, Caroline D.V.A. Bishop, Amy Jost, Elizabeth Crane, and Jennifer A. Punt. Isolation of T-Cell Subsets by Magnetic Cell Sorting (MACS), Luminita A. Stanciu and Ratko Djukanovic. Isolation of T-Cell Antigens by Retrovirus-Mediated Expression Cloning, Toshio Kitamura and Yoshihiro Morikawa. Flow Cytometric Analysis of Murine T Lymphocytes: A Practical Guide, Melanie S. Vacchio and Elizabeth W. Shores. Production and Characterization of Immature Murine T-Lymphoma Cell Lines, Ellen R. Richie and Dawn A. Walker. Production and Characterization of T-Cell Hybridomas, Janice White, John Kappler, and Philippa Marrack. Part III. Overview: Something Happens. A Brief History of TCR Signal Transduction, John J. O'Shea. Part IV. T Cell Activation. Measurement of Phosphatidylinositol (PI) Hydrolysis in Activated T Lymphocytes, Seth Berney and T. Prescott Atkinson. Fluorescence Polarization as an Early Measure of T-Lymphocyte Stimulation, Mordechai Deutsch, Naomi Zurgil, Menachem Kaufman, and Gideon Berke. Measurement of Lymphoproliferation at the Single-Cell Level by Flow Cytometry, Hans Gaines and Gunnel Biberfeld. Digital Image Analysis of Lymphocyte Activation, Bradley W. McIntyre, T. Kent Teague, Darren G. Woodside, and David K. Wooten. Induction of T-Cell Signaling by Immobilized Integrin Ligands, Jean Maguire van Seventer and Gijs A.van Seventer. Cytotoxic T-Cell Adherence Assay (CAA), Graham R. Leggatt and Jay A. Berzofsky. Detecting Ubiquitinated T-Cell Antigen Receptor Subunits by Immunoblotting, Allan M. Weissman. Determination of CD45 Tyrosine Phosphatase Activity in T Lymphocytes, David H. W. Ng, Jackie Felberg, and Pauline Johnson. Measurement of Protein Tyrosine Phosphorylation in T-Cell Subsets by Flow Cytometry, Dariush Farahi Far and Bernard Rossi. Biochemical Analysis of Activated T Lymphocytes: Protein Phosphorylation and Ras, ERK, and JNK Activation, Patrick E. Fields and Thomas F. Gajewski. Activation of Heterotrimeric GTP-Binding Proteins Upon TCR/CD3 Engagement, Constantine D. Tsoukas, Jack Stanners, and Keith A. Ching. Generation of Human T Helper 1 and T Helper 2 Subsets from Peripheral Blood-Derived Naive CD4+ T Cells, Ellen M. Palmer, Roshanak Tolouei Semnani, Bradford L. McRae, and Gijs A. van Seventer. Transcriptional Expression Analysis of T-Cell Activation by Multiplex Messenger Assay (MMA), Karine Bernard, Samuel Granjeaud, Genevieve Victorero, and Catherine Nguyen. Index
Review :
The protocols themselves are highly detailed, and also refer to a notes section in each chapter which is used to impart tips and tricks which are usually essential to getting a technique to work well, but which do not often come over in the usual material and methods section of papers. Each of the chapters is also well referenced. The book would be invaluable for establishing a particular technique or set of techniques in an environment where there was little or no previous experience of such as as teaching aid for when new research staff or students come into a laboratory. - The Journal of Infection
