Neutrophil: Methods and Protocols

Description

Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. This volume provides a concise set of protocols for assessing basic neutrophil functions and investigating specialized areas in neutrophil biology. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. In the third edition of Neutrophils: Methods and Protocols, chapters retained from previous editions have been have been updated and include many new approaches. In addition, the Third Edition contains a number of new chapters that were not included in the first or second editions. Part I contains overviews of neutrophil biology, function, and disorders. Part II describes commonly used methods to isolate neutrophils from humans and other animal species. Part III details methods for investigating chemotaxis, transmigration, phagocytosis, and bactericidal activity. Part IV includes protocols that measure neutrophil apoptosis, calcium signal transduction, degranulation and detection of cytoplasmic granules, gene expression, transcription factors, and apoptosis. Part V provides multiple assays for measuring production of intracellular and/or extracellular reactive oxygen species, and a method that details use of the cell-free NADPH oxidase assay, an iconic assay for studies of the neutrophil NADPH oxidase. Part VI provides chapters that describe how to analyze formation and function of neutrophil extracellular traps. Neutrophil Methods and Protocols: Third 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. Features • 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 Readership Molecular and Cell Biologists, Immunologists, Infectious Disease Specialists, Microbiologists Front Matter ....Pages i-xvi Front Matter ....Pages 1-1 The Role of Neutrophils in the Immune System: An Overview (Harry L. Malech, Frank R. DeLeo, Mark T. Quinn)....Pages 3-10 Neutrophil Defects and Diagnosis Disorders of Neutrophil Function: An Overview (Mary C. Dinauer)....Pages 11-29 Front Matter ....Pages 31-31 Isolation of Human Neutrophils from Venous Blood (Silvie Kremserova, William M. Nauseef)....Pages 33-42 Isolation of Neutrophils from Nonhuman Species (Daniel W. Siemsen, Liliya N. Kirpotina, Natalia Malachowa, Igor A. Schepetkin, Adeline R. Porter, Benfang Lei et al.)....Pages 43-59 Isolation of Neutrophils from Larval Zebrafish and Their Transplantation into Recipient Larvae for Functional Studies (Hannah Darroch, Jonathan W. Astin, Christopher J. Hall)....Pages 61-75 Front Matter ....Pages 77-77 Analysis of Neutrophil Transmigration Through Epithelial Cell Monolayers (Liliya N. Kirpotina, Douglas J. Kominsky, Mark T. Quinn, Steve D. Swain)....Pages 79-91 Quantification of Chemotaxis or Respiratory Burst Using Ex Vivo Culture-Derived Murine Neutrophils (Klaudia Szymczak, Margery G. H. Pelletier, Peter C. W. Gaines)....Pages 93-106 Ex Vivo Human Neutrophil Swarming Against Live Microbial Targets (Alex Hopke, Daniel Irimia)....Pages 107-116 Microinjection and Micropipette-Controlled Phagocytosis Methods for Neutrophils (Maurice B. Hallett, Jennie S. Campbell, Iraj Laffafian, Sharon Dewitt)....Pages 117-125 Using Imaging Flow Cytometry to Quantify Neutrophil Phagocytosis (Asya Smirnov, Michael D. Solga, Joanne Lannigan, Alison K. Criss)....Pages 127-140 Visualization and Quantification of Phagocytosis by Neutrophils (Gaelen Guzman, Fikadu G. Tafesse)....Pages 141-148 Analysis of Neutrophil Bactericidal Activity (Nicholas J. Magon, Heather A. Parker, Louisa V. Ashby, Reuben J. Springer, Mark B. Hampton)....Pages 149-164 Front Matter ....Pages 165-165 Assessment of Neutrophil Apoptosis (Nicole D. Barth, Marc Vendrell, David A. Dorward, Adriano G. Rossi, Ian Dransfield)....Pages 167-190 Optical Methods for the Measurement and Manipulation of Cytosolic Calcium Signals in Neutrophils (Maurice B. Hallett, Rhiannon E. Roberts, Sharon Dewitt)....Pages 191-205 Labeling Acidic Compartments of Neutrophils with Cresyl Violet (Philip P. Ostrowski, Ziv Roth, Sergio Grinstein)....Pages 207-213 Neutrophil Degranulation of Azurophil and Specific Granules (Samia Bedouhène, Pham My-Chan Dang, Margarita Hurtado-Nedelec, Jamel El-Benna)....Pages 215-222 Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils (Katja Branitzki-Heinemann, Graham Brogden, Maren von Köckritz-Blickwede)....Pages 223-233 In Vitro Assay for Sensitive Determination of Human Blood PMN Responses (Noah Fine, William Khoury, Michael Glogauer)....Pages 235-241 Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Transcription Real-Time PCR (Nicola Tamassia, Marco A. Cassatella, Flavia Bazzoni)....Pages 243-260 Detection of Intact Transcription Factors in Human Neutrophils (Patrick P. McDonald, Richard D. Ye)....Pages 261-275 Genome-Scale Transcript Analyses of Human Neutrophils (Scott D. Kobayashi, Adeline R. Porter, Sarah L. Anzick, Dan E. Sturdevant, Frank R. DeLeo)....Pages 277-298 Front Matter ....Pages 299-299 Measurement of Respiratory Burst Products, Released or Retained, During Activation of Professional Phagocytes (Claes Dahlgren, Halla Björnsdottir, Martina Sundqvist, Karin Christenson, Johan Bylund)....Pages 301-324 Cell-Free NADPH Oxidase Activation Assays: A Triumph of Reductionism (Edgar Pick)....Pages 325-411 Front Matter ....Pages 413-413 Immunofluorescent Detection of NET Components in Paraffin-Embedded Tissue (Ulrike Abu-Abed, Volker Brinkmann)....Pages 415-424 Detection, Visualization, and Quantification of Neutrophil Extracellular Traps (NETs) and NET Markers (Nicole de Buhr, Maren von Köckritz-Blickwede)....Pages 425-442 Imaging of Neutrophils and Neutrophil Extracellular Traps (NETs) with Intravital (In Vivo) Microscopy (Iwona Cichon, Michal Santocki, Weronika Ortmann, Elzbieta Kolaczkowska)....Pages 443-466 Back Matter ....Pages 467-470