Neutrophil Methods and Protocols / Edition 1

by Mark T. Quinn, Frank R. DeLeo, Gary M. Bokoch

Overview

Neutrophil Methods and Prools provides a concise set of prools for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the prools is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Parts 1 & 2 provide an overview of neutrophils the methods most commonly used for isolating them from humans and other animal species. Part 3 details methods for investigating adhesion, chemotaxis, phagocytosis and bacterial activity. Part 4 presents prools measuring gene expression in neutrophils, and Part 5 concludes with assays for diagnosing the most common neutrophil disorders. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.

Product Details

ISBN-13:	9781617377792
Publisher:	Springer-Verlag New York, LLC
Publication date:	12/10/2010
Series:	Methods in Molecular Biology , #412
Edition description:	Softcover reprint of hardcover 1st ed. 2007
Pages:	537
Product dimensions:	5.98(w) x 9.02(h) x 0.24(d)

Table of Contents

Neutrophils

Methods and Prools

Edited by

Mark T. Quinn

Frank R. DeLeo

Gary M. Bokoch

Contents

Dedication i

Preface ii

Contributors v

PART I: NEUTROPHILS: AN OVERVIEW

1. The Role of Neutrophils in the Immune System: An Overview

Harry L. Malech

PART II: NEUTROPHIL ISOLATION AND SUBCELLULAR FRACTIONATION

2. Isolation of Human Neutrophils From Venous Blood

William M. Nauseef

3. Neutrophil Isolation From Non-Human Species

Daniel W. Siemsen, Igor A. Schepetkin, Lilia N. Kirpotina, Benfang Lei, and Mark T. Quinn

4. Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers

Lene Udby and Niels Borregaard

PART III. BIOCHEMISTRY, BIOLOGY AND SIGNAL TRANSDUCTION OF NEUTROPHILS

5. Rac and Rap GTPase Activation Assays

Ulla G. Knaus, Alison Bamberg, and Gary M. Bokoch

6. Measurement of Phospholipid Metabolism in Intact Neutrophils

Susan Sergeant and Linda C. McPhail

7. Analysis of Protein Phosphorylation in Human Neutrophils

Jamel El-Benna and Pham My-Chan Dang

8. Mitogen-Activated Protein Kinase Assays

Martine Torres

9. Transduction of Proteins into Intact Neutrophils

Tieming Zhao and Gary M. Bokoch

10. Optical Methods for the Measurement and Manipulation of Cytosolic Free Calcium in Neutrophils

Esther J. Hillson, Sharon Dewitt, and Maurice B. Hallett

11. Analysis of Electrophysiological Properties and Responses of Neutrophils

Deri Morgan and Thomas E. DeCoursey

12. Analysis of Neutrophil Apoptosis

Emma L. Taylor, Adriano G. Rossi, Ian Dransfield, and Simon P. Hart

PART IV. NEUTROPHIL ADHESION AND CHEMOTAXIS

13. Optical and Fluorescence Detection of Neutrophil Integrin Activation

Ulrich Y. Schaff, Melissa R. Sarantos, Harold Ting, and Scott I. Simon

14. Analysis of Neutrophil Polarization and Chemotaxis

Mary A. Lokuta, Paul A. Nuzzi, and Anna Huttenlocher

15. Quantifying and Localizing Actin Free Barbed Ends in Neutrophils

Michael Glogauer

16. Measurement of Neutrophil Adhesion Under Conditions Mimicking Blood Flow

Mark A. Jutila, Bruce Walcheck, Robert Bargatze, and Aiyappa Palecanda

17. Model Systems to Investigate Neutrophil Adhesion and Chemotaxis

Nancy A. Louis, Eric Campbell, and Sean P. Colgan

PART V. NEUTROPHIL PHAGOCYTOSIS AND BACTERICIDAL ACTIVITY

18. Immunofluorescence and Confocal Microscopy of Neutrophils

Lee-Ann H. Allen

19. Assessment of Phagosome Formation and Maturation by Fluorescence Microscopy

Benjamin E. Steinberg and Sergio Grinstein

20. Analysis of Neutrophil Membrane Traffic During Phagocytosis

Per Lönnbro, Pontus Nordenfelt, and Hans Tapper

21. Analysis of Neutrophil Bactericidal Activity

Jessie N. Green, Christine C. Winterbourn, and Mark B. Hampton

22. A Skin Chamber Technique as a Human Model for Studies of Aseptic Inflammatory Reactions

Per Follin and Claes Dahlgren

PART VI. NADPH OXIDASE AND PRODUCTION OF REACTIVE OXYGEN SPECIES

23. Measurement of Respiratory Burst Products Generated by Professional Phagocytes

Claes Dahlgren, Anna Karlsson, and Johan Bylund

24. The K-562 Cell Model for Analysis of Neutrophil NADPH Oxidase Function

Thomas L. Leto, Mark C. Lavigne, Neda Homoyounpour, Kristen Lekstrom, Gilda Linton, Harry L. Malech, and Isabelle de Mendez

25. Cell-free Assays: The Reductionist Approach to the Study of NADPH Oxidase Assembly

Shahar Molshanski-Mor, Ariel Mizrahi, Yelena Ugolev, Iris Dahan, Yevgeny Berdichevsky, and Edgar Pick

26. Immunoaffinity Purification of Human Phagocyte Flavocyhrome b and Analysis of Conformational Dynamics

Ross M. Taylor and Algirdas J. Jesaitis

PART VII. ANALYSIS OF NEUTROPHIL GENE EXPRESSION AND TRANSCRIPTION FACTORS

27. Genome-scale Transcript Analyses with Human Neutrophils

Scott D. Kobayashi, Dan E. Sturdevant, and Frank R. DeLeo

28. Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Transcription Real-Time PCR

Nicola Tamassia, Marco A. Cassatella, and Flavia Bazzoni

29. Detection of Intact Transcription Factors in Human Neutrophils

Patrick P. McDonald and Richard D. Ye

PART VIII. NEUTROPHIL DEFECTS AND DIAGNOSIS

30. Disorders of Neutrophil Function: An Overview

Mary C. Dinauer

31. Diagnostic Assays for Chronic Granulomatous Disease and Other Neutrophil Disorders

Houda Zghal Elloumi and Steven M. Holland

32. Diagnostic Assays for Myeloperoxidase Deficiency

William M. Nauseef

Editorial Reviews

Reviewer: Marion C. Cohen, PhD (SUNY Downstate Medical Center)
Description: This volume in the Methods in Molecular Biology series is a comprehensive collection of detailed protocols currently used to study neutrophil biology. In addition chapters provide background for the assays that follow.
Purpose: According to the editor, the book provides a set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and directions for diagnostic assays of common neutrophil disorders. While one laboratory may not make use of all, or even many, of the protocols, they are useful to have in one volume as a reference. Moreover, they are more detailed in terms of troubleshooting than one is likely to find in many other methods books.
Audience: The book is targeted at basic researchers involved in the study of neutrophil function and those doing clinical research on how neutrophils function in specific diseases. The chapter authors are authorities in their fields.
Features: The eight sections include an overview, a section on neutrophil isolation, sections that focus on groups of various neutrophil functions, a section on gene expression and transcription factors, and a section on neutrophil defects and diagnosis. The result is a small encyclopedia on the neutrophil. The detailed experimental protocols are very helpful. The use of "Notes" to better explain some issues is a characteristic of the books in this series and provides a way of dealing with small details that rarely show up in the methods section of a paper. Likewise, the overview chapters are a good addition. Most of the illustrations are black and white and those that are also presented in color are grouped together in one section of the book.
Assessment: This is a carefully compiled collection of protocols that allow for study of many aspects of the neutrophil. The experimental design is described in considerable detail and the overviews are helpful. Since there are more than 400 books in the series, one must conclude that this is a format that works well.

Doody's Review Service

From the reviews:

"...a carefully compiled collection of prools that allow for study of many aspects of the neutrophil." -Doody's Book Review, Weighted Numerical Score:85-3 Stars

"This excellent book is one of the latest in the ‘Methods in Molecular Biology’ series … . it comes beautifully presented and bound in a hardback cover and would grace any bookshelf. I would urge all self-respecting laboratories working with neutrophils – whether in research or diagnostics – to purchase one immediately. … I was impressed by this book’s comprehensive provision of prools, addressing all aspects of neutrophil function and physiology, and including an excellent section on diagnosis of both inherited and non-inherited neutrophil disorders." (Lynn Morgan, Immunology News, May, 2008)

From the Publisher