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per Medici e Operatori Sanitari
per Medici e Operatori Sanitari
LIBRERIA STUDIUM
Neutrophil Methods and Protocols, 2nd ed
Series: Methods in Molecular Biology, Vol. 1124
Quinn, DeLeo
Editore
Humana Press - Springer
Anno
2014
Pagine
551
ISBN
9781627038447
125,00 €
- Provides neutrophil protocols that are broadly applicable to leukocyte biology
- Includes protocols for isolation of neutrophils from humans and animal species
- Presents an interesting and informative overview of the field of neutrophil biology
- Facilitates a better understanding of the role of neutrophils in host defense
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.
Contents
Preface .................................................................................................................... vii
Contributors .................................................................................................................... xiii
PART I NEUTROPHILS: AN OVERVIEW
1 The Role of Neutrophils in the Immune System: An Overview ......................... 3
Harry L. Malech, Frank R. DeLeo, and Mark T. Quinn
PART II NEUTROPHIL ISOLATION AND SUBCELLULAR FRACTIONATION
2 Isolation of Human Neutrophils from Venous Blood ........................................ 13
William M. Nauseef
3 Neutrophil Isolation from Nonhuman Species .................................................. 19
Daniel W. Siemsen, Natalia Malachowa, Igor A. Schepetkin,
Adeline R. Whitney, Liliya N. Kirpotina, Benfang Lei, Frank
R. DeLeo, and Mark T. Quinn
4 Collection of In Vivo Transmigrated Neutrophils from Human Skin ................. 39
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 .................................................................. 53
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 ................................................. 79
Richard T. Jennings and Ulla G. Knaus
7 Measurement of Phospholipid Metabolism in Intact Neutrophils ...................... 89
Susan Sergeant and Linda C. McPhail
8 Optical Methods for the Measurement and Manipulation
of Cytosolic Calcium Signals in Neutrophils ..................................................... 107
Maurice B. Hallett, Maha Al-Jumaa, and Sharon Dewitt
9 Analysis of Electrophysiological Properties and Responses of Neutrophils ......... 121
Deri Morgan and Thomas E. DeCoursey
10 Assessment of Neutrophil Apoptosis ................................................................. 159
David A. Dorward, Adriano G. Rossi, Ian Dransfield,
and Christopher D. Lucas
x
11 Microinjection Methods for Neutrophils .......................................................... 181
Iraj Laffafian, Kimberly J. Lewis, K.Benjamin Masterman,
and Maurice B. Hallett
12 Generation of Functionally Mature Neutrophils from Induced
Pluripotent Stem Cells ...................................................................................... 189
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 ......................................... 209
Richard T. Jennings and Ulla G. Knaus
14 Spinning Disk Confocal Imaging of Neutrophil Migration in Zebrafish ............ 219
Pui-ying Lam, Robert S. Fischer, William D. Shin, Clare M. Waterman,
and Anna Huttenlocher
15 Detection of Bidirectional Signaling During Integrin Activation
and Neutrophil Adhesion ................................................................................. 235
Stuart M. Altman, Neha Dixit, and Scott I. Simon
PART V NEUTROPHIL PHAGOCYTOSIS AND BACTERICIDAL ACTIVITY
16 Immunofluorescence and Confocal Microscopy of Neutrophils ......................... 251
Lee-Ann H. Allen
17 Expression of Genetically Encoded Fluorescent Probes
to Monitor Phospholipid Dynamics in Live Neutrophils ................................... 269
Benjamin E. Steinberg, Marco A.O. Magalhaes, and Sergio Grinstein
18 Quantitative Assessment of Neutrophil Phagocytosis
Using Flow Cytometry ..................................................................................... 279
Pontus Nordenfelt
19 Analysis of Neutrophil Bactericidal Activity ....................................................... 291
Heather A. Parker, Nicholas J. Magon, Jessie N. Green,
Mark B. Hampton, and Christine C. Winterbourn
20 Induction and Quantification of Neutrophil Extracellular Traps ........................ 307
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 ............................... 321
Johan Bylund, Halla Björnsdottir, Martina Sundqvist,
Anna Karlsson, and Claes Dahlgren
22 Cell-Free NADPH Oxidase Activation Assays: “In Vitro Veritas” ...................... 339
Edgar Pick
Contents
xi
23 Assessment of Priming of the Human Neutrophil Respiratory Burst ................. 405
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 .............................................................................................. 413
Marcia Riesselman and Algirdas J. Jesaitis
25 Evaluation of p47phox Phosphorylation in Human Neutrophils
Using Phospho-Specific Antibodies .................................................................. 427
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 ............................ 437
Scott D. Kobayashi, Daniel E. Sturdevant, and Frank R. DeLeo
27 Fast and Accurate Quantitative Analysis of Cytokine
Gene Expression in Human Neutrophils ........................................................... 451
Nicola Tamassia, Marco A. Cassatella, and Flavia Bazzoni
28 High-Purity Neutrophil Isolation from Human Peripheral Blood
and Saliva for Transcriptome Analysis ................................................................ 469
Flavia S. Lakschevitz and Michael Glogauer
29 Detection of Intact Transcription Factors in Human Neutrophils ...................... 485
Patrick P. McDonald and Richard D. Ye
PART VIII NEUTROPHIL DEFECTS AND DIAGNOSIS
30 Disorders of Neutrophil Function: An Overview ............................................... 501
Mary C. Dinauer
31 Diagnostic Assays for Chronic Granulomatous Disease
and Other Neutrophil Disorders ....................................................................... 517
Houda Zghal Elloumi and Steven M. Holland
32 Diagnostic Assays for Myeloperoxidase and Myeloperoxidase Deficiency ........... 537
William M. Nauseef
Index ...................................................
