Human Airway Inflammation: Sampling Techniques and Analytical Protocols

ISBN-13: 9781489943729 / Angielski / Miękka / 2013 / 440 str.

Duncan F. Rogers; Louise E. Donnelly

Opis

Rubor (redness), tumor (swelling), calor (heat), and dolor (pain) are the classical signs of inflammation. These features are obvious in the skin, where injury or disease causes flare, wheal, and painful burning sensations. Vasodi- tation underlies the flare and heat, plasma exudation the swelling, and acti- tion of sensory nerves relays pain. In chronic conditions, skin biopsies show inflammatory cell infiltrate. Inflammation is not unique to the skin and contr- utes to disease and repair processes in other organ systems in the body. From the viewpoint of this volume, lung inflammation is now recognized as central to the pathophysiology of a number of severe respiratory conditions, the two most common being asthma and chronic obstructive pulmonary disease (COPD). In asthma, and to a lesser extent COPD, there is evidence of vasodilatation, with congestion of blood vessels accompanied by reddening of the airway mucosa, and of plasma exudation, leading to swelling of the airway wall. Similarly, although less pronounced than in the skin, there is evidence of pain, for example, the - pleasant chest sensations associated with asthma attacks. Understanding the pat- genesis of airway inflammation will enable rational design of drugs to effectively treat conditions such as asthma and COPD. However, whereas immediate access to the skin facilitates investigation of disease processes, the lung, although "open to atmosphere," is much less accessible. Consequently, the investigation of lung inflammation is usually indirect. Thus, a wide variety of research techniques are used.

Recenzje

"Comprehensive and highly practical, the methods presented in this volume provide today's basic and clinical researchers all the major techniques for investigating airway inflammation, and powerfully illuminate many novel targets for emerging drugs." - Journal of Investigational Allergology and Clinical Immunology

Spis treści

Airway Inflammation and Remodeling in Asthma: Current Concepts Stephen T. Holgate Part I. Sample Collection: Invasive Techniques Biopsy Techniques: Optimization for Collection and Preservatio Marina Saetta and Graziella Turato Bronchoalveolar Lavage (BAL): Critical Evaluation of Techniques Christopher Ward and E. Haydn Walters Nasal Secretions and Exudations: Collection and Approaches to Analysis Lennard Greiff, Morgan Andersson, and Carl G.A. Persson Part II. Sample Collection: Noninvasive Techniques Induced Sputum: Selection Method Margaret M. Kelly, Ann Efthimiadis, and Frederick E. Hargreave Induced Sputum: Whole Sample Vera M. Keatings and Julia A. Nightingale Measurement of Exhaled Nitric Oxide and Carbon Monoxide Sergei A. Kharitonov Measurement of Exhaled Hydrocarbons Paolo Paredi, Sergei A. Kharitonov, and Peter J. Barnes Breath Condensate as a Vehicle for Collection of Inflammatory Mediators, Especially Hydrogen Peroxide Johan C. de Jongste, Rijn Jöbsis, and H. Rolien Raatgeep Part III. Cell Isolation and Culture Airway Epithelial Cells (Primaries vs Cell Lines) Louise E. Donnelly Isolation and Culture of Human Alveolar Type II Pneumocytes Ian R. Witherden and Teresa D. Tetley Pulmonary Artery Endothelial Cells Joachim Seybold and Norbert Suttorp Isolation and Culture of Human Airway Smooth Muscle Cells Aili L. Lazaar and Reynold A. Panettieri, Jr Isolation and Purification of Human Mast Cells and Basophils Bernhard F. Gibbs and Madeleine Ennis Neutrophils: Collection, Separation, and Activation Sarah V. Culpitt Macrophages: Identification, Separation, and Function Leonard W. Poulter and C.M. Burke Collection, Separation, and Activation of Human T Lymphocytes YannisSotsios and Stephen G. Ward Eosinophils: Collection, Separation, and Activation Leo Koenderman, Jan van der Linden, Laurien Ulfman, and Paul Coffer Part IV. Gene Expression/Regulation Analysis of Gene Expression: Reverse Transcription-Polymerase Chain Reaction El-Bdaoui Haddad and Jonathan Roussell Analysis of Gene Expression: Northern Blotting Jonathan Roussell and El-Bdaoui Haddad The Technique of In Situ Hybridization Sun Ying and A. Barry Kay Measurement of Airway Mucin Gene Expression Kelly Pritchard, Alinka K. Smith, and Duncan F. Rogers Transcription Factors and Inflammatory Gene Regulation: Strategic Approaches Robert Newton and Karl J. Staples Part V. Mediators: Expression and Function Inflammatory Mediators in Spontaneously Produced Sputum Adam Hill, Simon Gompertz, Darren Bayley, and Robert Stockley Flow Cytometry: Measurement of Leukocyte Receptor Expression and Function Adele Hartnell Measurement of Granulocyte Pharmacodynamics in Whole Blood by Flow Cytometry Shannon A. Bryan, Margaret J. Leckie, Gavin Jenkins, Peter J. Barnes, Timothy J. Williams, Ian Sabroe, and Trevor T. Hansel Current In Vitro Models of Leukocyte Migration: Methods and Interpretation Jennifer R. Allport, Guillermo Garcia-Cardeña, Yaw-Chyn Lim, and Francis W. Luscinskas Tracing Intracellular Mediator Storage and Mobilization in Eosinophils Salahaddin Mahmudi-Azer, Paige Lacey, and Redwan Moqbel Measurement of Metalloproteinases Steven D. Shapiro, Diane Kelley and Dale Kobayashi Immunocytochemistry and Immunohistochemistry Jochen Springer and Axel Fischer The Measurement of Cysteinyl Leukotrienes in Urine Jay Y. Wescott and Sally E. Wenzel Quantitative Analysis of Cyclooxygenase Metabolites of Arachidonic Acid Ryszard

Blurb

Airway inflammation is now recognized as central to the pathophysiology of several major respiratory conditions-the two most common being asthma and chronic obstructive pulmonary disease (COPD). In Human Airway Inflammation: Sampling Techniques and Analytical Protocols, experienced basic and clinical investigators describe a wide range of readily reproducible methods for sampling, isolating, and culturing all of the major inflammatory cells involved in the pathophysiology of asthma and COPD. The collection techniques detailed here involve biopsy and sampling of airway liquids such as sputum and exhaled gases such as nitric oxide. Also provided is a full range of methods for the isolation and characterization of cells and for the measurement of many of the major inflammatory markers, along with protocols for measuring the inflammatory mediators and enzymes released during lung inflammation. Each user-friendly method contains all the essential step-by-step elements the researcher must navigate from sample collection to analysis, and includes notes on how to avoid problems. Among the more advanced techniques are the measurement of exhaled hydrocarbons and F2 isoprostanes, granulocyte pharmacodynamics in whole blood measured by flow cytometry, and tracing mediator trafficking in eosinophils using confocal microscopy.
Comprehensive and highly practical, the methods presented in Human Airway Inflammation: Sampling Techniques and Analytical Protocols provide today's basic and clinical researchers with all the major techniques for investigating airway inflammation, and powerfully illuminate many novel targets for emerging drugs.