T cell protocols


    • De Libero, Gennaro


T cell protocols

edited by Gennaro De Libero

(Methods in molecular biology / John M. Walker, series editor, 514)(Springer protocols)

Humana Press, c2009

2nd ed

  • : hbk

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Includes bibliographical references and index



This book is a collection of protocols, to provide novel techniques for the study of the biology of T lymphocytes. The methods described in this book do not cover all of the techniques currently used to study T cell-mediated immune responses for the simple reason that T cell immunology is probably the immunological discipline which can be investigated with the widest variety of approaches. Thechoiceofchapterswasmadetakingintoaccounttwopoints:First,manyofthe techniquesthathavebeen usedfor sometimehave beenupgraded duringthe pastfew years given the greater availability of a variety of products (i.e. cytokines, chemokines, monoclonal antibodies), of refined technical devices (i.e. novel cell culture and cell analysis equipments), and the development of novel instrumentation (i.e. multipa- metric flow cytometers, confocal microscopes). Therefore, in several chapters ``old techniques'', which remain fundamental to T cell immunology, are described in their ``modern'' versions. Secondly, the technical advancement has generated the possibility to establish novel assays to investigate T cell physiology. This is reflected in the chapters which describe the protocols that allow use of these modern approaches. The preparation of this book has required participation of several scientists, all leading experts in their respective fields. Without their enthusiastic participation, this work would not have been possible. Therefore, I thank all authors for their contri- tionsandacceptallcriticismformissingparts,orinformationordetails,forwhichonly I am responsible. I hope these protocols will be useful for young investigators who approach for the first time the complex field of immunology and for those more experienced scientists who look for concise and efficacious descriptions of novel methods.


Chapter 1: Analysis of Frequency and Phenotype of Antigen-Specific T Cells Angus Stock and Vincenzo Cerundolo Chapter 2: B Cell Helper Assays Sergio Abrignani, Elena Tonti, Giulia Casorati, and Paolo Dellabona Chapter 3: Trans-Kingdom RNA Interference (tkRNAi): A Novel Method to Induce Therapeutic Gene Silencing Thu A. Nguyen and Johannes H. Fruehauf Chapter 4: Flow Cytometry and Cell Activation Sonia Gavasso Chapter 5: Investigating T Cells by Polychromatic Flow Cytometry Enrico Lugli, Leonarda Troiano, and Andrea Cossarizza Chapter 6: Generation of Human T Cell Clones Sabrina Mariotti and Roberto Nisini Chapter 7: Limiting Dilution Analysis of Antigen-Specific T cells Jorge Carneiro, Lurdes Duarte, and Elisabetta Padovan Chapter 8: T Cell Epitope-Mapping by Cytokine Gene Expression Assay Maurizio Provengano and Giulio C. Spagnoli Chapter 9: Cytokine Multiplex Immunoassay: Methodology and (Clinical) Applications Wilco de Jager, Berent Prakken, and Ger T. Rijkers Chapter 10: Purification of the T Cell Antigen Receptor and Analysis by Blue-Native PAGE Mahima Swamy and Wolfgang W.A. Schamel Chapter 11: Non-Replicating Recombinant Vaccinia Virus Expressing CD80 to Enhance T-Cell Stimulation Paul Zajac

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