Handbook of Elispot: Methods and Protocols

ISBN-13: 9781617793240 / Angielski / Twarda / 2011 / 261 str.

Alexander E. Kalyuzhny

Opis

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.

Recenzje

From the reviews of the second edition:

"In this second edition ... the reader is introduced not only to the basic concept behind the technique but also to many different applications in common use. Importantly, the book serves as a reference guide with lots of trouble shooting and discussion of common problems. ... This text would be key reading material for any laboratory planning to set up this technique. ... this text is comprehensive and detailed with a good use of raw data and formula to make following the protocols simple." (Hayley Evans, Immunology News, November, 2012)

Spis treści

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 Analysis

11. 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 IFNγ 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 Analysis

11. 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 Analysis

11. 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 Analysis

11. 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 Analysis

11. 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 IFNγ 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-γ 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 Analysis

11. 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 IFNγ 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-γ 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