Neutrophil Methods and Protocols

ISBN-13: 9781627038447 / Angielski / Twarda / 2014 / 551 str.

Mark T. Quinn; Frank R. DeLeo

Opis

Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. Neutrophil Methods and Protocols, Second Edition provides a concise set of protocols written by leading researchers in the field for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step diagnostic assays of common neutrophil disorders. Topics covered include an overview of neutrophils and their role in host defense and inflammation; methods most commonly used for isolating neutrophils from humans and other animal species; procedures for subcellular fractionation of human neutrophils, analysis of in vivo transmigrated neutrophils, generation of mature neutrophils from induced pluripotent stem cells and analysis of neutrophil gene expression; methods for investigating priming, oxidant production, phagocytosis, bactericidal activity and extracellular trap formation and protocols for investigating neutrophil adhesion, chemotaxis and outside-in signaling via integrins. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and easily accessible, Neutrophil Methods and Protocols, Second Edition is a comprehensive source for detailed explanations and applications of the most modern methodological advances in neutrophil biology. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.

Spis treści

Part I. Neutrophils: an Overview

 

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

Harry L. Malech, Frank R. DeLeo, and Mark T. Quinn

 

 

Part II. Neutrophil Isolation and Subcellular Fractionation

 

2. Isolation of Human Neutrophils

William M. Nauseef

 

3. Neutrophil Isolation from Non-Human Species

Daniel W. Siemsen, Natalia Malachowa, Igor A. Schepetkin, Adeline R. Whitney, Lilya N. Kirpotina, Benfang Lei, Frank R. DeLeo, and Mark T. Quinn

 

4. Collection of In vivo Transmigrated Neutrophils from Human Skin

Karin Christenson, Lena Björkman, Lisa Davidsson, Anna Karlsson, Per Follin, Claes Dahlgren, and Johan Bylund

 

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

Stine Novrup Clemmensen, Lene Udby, and Niels Borregaard

 

           

Part III. Biochemistry, Biology and Signal Transduction of Neutrophils

 

6. Rho Family and Rap GTPase Activation Assays

Richard T. Jennings and Ulla G. Knaus

 

7. Measurement of Phospholipid Metabolism in Intact Neutrophils

Susan Sergeant and Linda C. McPhail

 

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

Maurice B. Hallett , Maha Al-Jumaa, and  Sharon Dewitt

 

9. Analysis of Electrophysiological Properties and Responses of Neutrophils

Deri Morgan and Thomas E. DeCoursey

 

10. Analysis of Neutrophil Apoptosis

David A. Dorward, Adriano G. Rossi, Ian Dransfield, and Christopher D. Lucas

 

11. Microinjection Methods for Neutrophils

Iraj Laffafian, Kimberly J. Lewis, K. Benjamin Masterman, and Maurice B. Hallett

 

12. Generation of Functionally Mature Neutrophils from Induced Pluripotent Stem Cells

Colin L. Sweeney, Randall K. Merling, Uimook Choi, Debra Long Priel, Douglas B. Kuhns, Hongmei Wang, and Harry L. Malech

 

 

Part IV. Neutrophil Adhesion and Chemotaxis

 

13. Neutrophil Migration Through Extracellular Matrix

Richard T. Jennings and Ulla G. Knaus

 

14. Spinning Disk Confocal Imaging of Neutrophil Migration in Zebrafish

Pui-ying Lam, Robert S. Fischer, William D. Shin, Clare M. Waterman, and Anna Huttenlocher

 

15. Detection of Bi-Directional Signalilng During Integrin Activation and In Neutrophil Adhesion

Stuart M. Altman, Neha Dixit, and Scott I. Simon

 

 

Part V. Neutrophil Phagocytosis and Bactericidal Activity

 

16. Immunofluorescence and Confocal Microscopy of Neutrophils

Lee-Ann H. Allen

 

17. Expression of Genetically-Encoded Fluorescent Probes to Monitor Phospholipid Dynamics in Live Neutrophils

Benjamin E. Steinberg, Marco A.O. Magalhaes, and Sergio Grinstein

 

18. Quantitative Assessment of Neutrophil Phagocytosis Using Flow Cytometry   

Pontus Nordenfelt

 

19. Analysis of Neutrophil Bactericidal Activity

Heather A. Parker, Nicholas J. Magon, Jessie N. Green, Mark B. Hampton, and Christine C. Winterbourn

 

20. Induction and Quantification of Neutrophil Extracellular Traps

Alejandro Sanchez Gonzalez, Bart W. Bardoel, Christopher J. Harbort, and Arturo Zychlinsky

 

 

Part VI. NADPH Oxidase and Production of Reactive Oxygen Species

 

21. Measurement of Respiratory Burst Products, Released or Retained, During Activation of Professional Phagocytes

Johan Bylund, Halla Björnsdottir, Martina Sundqvist, Anna Karlsson, and Claes Dahlgren

 

22. Cell-free NADPH Oxidase Assays: “In vitro Veritas”

Edgar Pick

 

23. Assessment of Priming of the Human Neutrophil Respiratory Burst

Margarita Hurtado-Nedelec, Karama Makni-Maalej, Marie-Anne Gougerot-Pocidalo, Pham My-Chan Dang, and Jamel El-Benna

 

24. Affinity Purification and Reconstitution of Human Phagocyte Flavocytochrome b for Detection of Conformational Dynamics in the Membrane

Marcia Riesselman and Algirdas J. Jesaitis

 

25. Evaluation of p47phox Phosphorylation in Human Neutrophils Using Phospho-Specific Antibodies

Sahra Amel Belambri, Pham My-Chan Dang, and Jamel El-Benna

 

 

Part VII. Analysis of Neutrophil Gene Expression and Transcription Factors

 

26. Genome-Scale Transcript Analyses with Human Neutrophils

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

 

27. Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils

Nicola Tamassia, Marco A. Cassatella, and Flavia Bazzoni

 

28. High Purity Neutrophil Isolation from Human Peripheral Blood and Saliva for Transcriptome Analysis

Flavia S. Lakschevitz and Michael Glogauer

 

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 and Myeloperoxidase Deficiency

William M. Nauseef