Handbook of ELISPOT

Enzyme-linked immunospot assay (ELISPOT) has been known for some time as a unique state-of-the-art technique for studying the cytokine-secreting activity of immune system cells, and it appears to be one of the fast growing applications in biomedical research, becoming an indispensable tool in vaccine development, HIV research, transplantation studies, and cancer and allergy research.  The second edition of Handbook of ELISPOT: Methods and Protocols, only the second book in the field which is entirely dedicated to ELISPOT assay, shares the detailed techniques that have been developed since the release of the popular first edition.  Straight from the labs of seasoned experts, this book covers setting and performing ELISPOT assays, ELISPOT for veterinary research, advanced ELISPOT techniques, image and data analysis, as well as vaccine development and diagnostics.  Written in the highly successful Methods in Molecular Biology- series format, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.  Authoritative and cutting-edge, Handbook of ELISPOT: Methods and Protocols, Second Edition serves as a compilation of a technical reference and a troubleshooting guide for researchers, both experienced and novice, worldwide in order to advance the usage of this key tool.

Part I: Setting and Performing ELISPOT Assay 1. Unique Strengths of ELISPOT for T Cell Diagnostics            Paul V. Lehmann and Wenji Zhang 2. The Impact of Harmonization on ELISPOT Assay Performance 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Sylvia Janetzki and Cedrik M. Britten Part II: ELISPOT for Veterinary Research 3. Equine ELISPOT Assay to Study Secretion of IFN- and IL-4 from Peripheral Blood Mononuclear Cells 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Jodi Hagen, Chris Hartnett, J.P. Houchins, Steeve Giguère, and Alexander E. Kalyuzhny 4. Utilization of Feline ELISPOT for Mapping Vaccine Epitopes 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Jeffrey R. Abbott, Ruiyu Pu, James K. Coleman, and Janet K. Yamamoto 5. Analyzing Cellular Immunity to AAV in a Canine Model Using ELISPOT Assay 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Zejing Wang, Rainer Storb, Stephen J. Tapscott, and Stanley Riddell Part III: Advanced ELISPOT Techniques 6. Dual and Triple Color Fluorospot 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Niklas Ahlborg and Bernt Axelsson            Alexey Y. Karulin and Paul V. Lehmann 12. Mathematical Algorithms for Automatic Search, Recognition, and Detection of Spots in ELISPOT Assay            Sergey S. Zadorozhny and Nikolai N. Martynov 13. Objective, User-Independent ELISPOT Data Analysis Based on Scientifically Validated Principles            Wenji Zhang and Paul V. Lehmann 14. Statistical Analysis of ELISPOT Assays            Marcus Dittrich and Paul V. Lehmann 15. Response Determination Criteria for ELISPOT: Toward a Standard that Can Be Applied Across Laboratories            Zoe Moodie, Leah Price, Sylvia Janetzki, and Cedrik M. Britten Part V: ELISPOT Assay for Vaccine Development and Diagnostics 16. Detection of Vaccinia Virus-Specific IFN- and IL-10 Secretion from Human PBMCs and CD8+ T Cells by ELISPOT            Benjamin J. Umlauf, Norman A. Pinsky, Inna G. Ovsyannikova, and Gregory A. Poland 17. ELISPOT Assays to Enumerate Bovine IFN-- Secreting Cells for the Development of Novel Vaccines Against Bovine Tuberculosis            Martin Vordermeier and Adam O. Whelan 18. IL-7 Addition Increases Spot Size and Number as Measured by T-SPOT.TB®            Marsha L. Feske, Miguel Medina, Edward A. Graviss, and Dorothy E. Lewis 19. Overview of Membranes and Membrane Plates Used in Research and Diagnostic ELISPOT Assays            Alan J. Weiss 7. ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 8. ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny 9. ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion            Jun-Seop Shin and Chung-Gyu Park 10. Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell            Jodi Hagen, Jeffrey P. Houchins, and Alexander E. Kalyuzhny Part IV. Image and Data Analysis11. How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells- Secretory Activity            Alexey Y. Karulin and Paul V. Lehmann 12. Mathematical Algorithms for Automatic Search, Recognition, and Detection of Spots in ELISPOT Assay            Sergey S. Zadorozhny and Nikolai N. Martynov 13. Objective, User-Independent ELISPOT Data Analysis Based on Scientifically Validated Principles            Wenji Zhang and Paul V. Lehmann 14. Statistical Analysis of ELISPOT Assays            Marcus Dittrich and Paul V. Lehmann 15. Response Determination Criteria for ELISPOT: Toward a Standard that Can Be Applied Across Laboratories            Zoe Moodie, Leah Price, Sylvia Janetzki, and Cedrik M. Britten Part V: ELISPOT Assay for Vaccine Development and Diagnostics 16. Detection of Vaccinia Virus-Specific IFN- and IL-10 Secretion from Human PBMCs and CD8+ T Cells by ELISPOT            Benjamin J. Umlauf, Norman A. Pinsky, Inna G. Ovsyannikova, and Gregory A. Poland 17. ELISPOT Assays to Enumerate Bovine IFN-- Secreting Cells for the Development of Novel Vaccines Against Bovine Tuberculosis            Martin Vordermeier and Adam O. Whelan 18. IL-7 Addition Increases Spot Size and Number as Measured by T-SPOT.TB®            Marsha L. Feske, Miguel Medina, Edward A. Graviss, and Dorothy E. Lewis 19. Overview of Membranes and Membrane Plates Used in Research and Diagnostic ELISPOT Assays            Alan J. Weiss