T Cell Protocols

1. Front Matter

   Pages i-xi

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2. Analysis of Frequency and Phenotype of Antigen-Specific T Cells

   • Angus Stock, Vincenzo Cerundolo

   Pages 1-14

3. B Cell Helper Assays

   • Sergio Abrignani, Elena Tonti, Giulia Casorati, Paolo Dellabona

   Pages 15-26

4. transkingdom RNA Interference (tkRNAi): A Novel Method to Induce Therapeutic Gene Silencing

   • Thu A. Nguyen, Johannes H. Fruehauf

   Pages 27-34

5. Flow Cytometry and Cell Activation

   • Sonia Gavasso

   Pages 35-46

6. Investigating T Cells by Polychromatic Flow Cytometry

   • Enrico Lugli, Leonarda Troiano, Andrea Cossarizza

   Pages 47-63

7. Generation of Human T Cell Clones

   • Sabrina Mariotti, Roberto Nisini

   Pages 65-93

8. Limiting Dilution Analysis of Antigen-Specific T Cells

   • Jorge Carneiro, Lurdes Duarte, Elisabetta Padovan

   Pages 95-105

9. T Cell Epitope-Mapping by Cytokine Gene Expression Assay

   • Maurizio Provenzano, Giulio C. Spagnoli

   Pages 107-118

10. Cytokine Multiplex Immunoassay: Methodology and (Clinical) Applications

    • Wilco de Jager, Berent Prakken, Ger T. Rijkers

    Pages 119-133

11. Purification of the T Cell Antigen Receptor and Analysis by Blue-Native PAGE

    • Mahima Swamy, Wolfgang W.A. Schamel

    Pages 135-150

12. Non-Replicating Recombinant Vaccinia Virus Expressing CD80 to Enhance T-Cell Stimulation

    • Paul Zajac

    Pages 151-162

13. Back Matter

    Pages 163-165

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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 efficaciousdescriptions of novel methods.

• Flow cytometry
• Immune responses
• Polyacrylamid-Gelelektrophorese
• RNA
• T cell immunology
• T lymphocytes
• gene expression
• genes
• molecular biology
• Cell culture

From the reviews of the second edition:

"A great collection of protocols, some totally new and some upgraded … to study the biology of this fascinating cell is wellcome by the scientific community. … Thanks to the generation of … PFC, is now at the hands of those scientists that want to advance our understanding of T cells biology. … the book will be of enourmous help both to beginners and to experienced scientists is well underlining the esprit of the book, as I showed with the three examples illustrated." (CarloAlberto Redi, European Journal of Histochemistry, October, 2009)

• University of Basel, Switzerland

  Gennaro Libero

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