Human Airway Inflammation: Sampling Techniques and Analytical Protocols

435 Pages·2001·2.863 MB·English

by  Stephen T. Holgate (auth.), Duncan F. Rogers, Louise E. Donnelly (eds.)

Human Airway Inflammation
M E T H O D S I N M O L E C U L A R M E D I C I N ETM
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M E T H O D S I N M O L E C U L A R M E D I C I N ETM
Human Airway
Inflammation
Sampling Techniques
and Analytical Protocols
Edited by
Duncan F. Rogers
and
Louise E. Donnelly
National Heart and Lung Institute, Imperial College, London, UK
Humana Press Totowa, New Jersey
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Cover illustrations: Background is Fig. 4 from Chapter 10, “Airway Epithelial Cells (Primaries vs. Cell
Lines)” by Louis E. Donnelly; foreground is Fig. 1 from Chapter 13, “Isolation and Culture of Human Airway
Smooth Muscle Cells” by Aili L. Lazaar and Reynold A. Panettieri, Jr.
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Main entry under title:
Methods in molecular medicine™.
Human airway inflammation: smapling techniques and analytical protocols / edited by Duncan F. Rogers
and Louise E. Donnelly.
p.;cm.—(Methods in molecular medicine; 56)
Includes bibliographic references and index.
ISBN 0-89603-923-4 (alk. paper)
1. Airway (Medicine)–Inflammation–Diagnosis. 2. Asthma–Diagnosis. 3. Lungs–Diseases,
Obstructive Diagnosis. 4. Molecular diagnosis. I. Rogers, D. F. (Duncan F.), 1953–II.
Donnelly, Louise E. III. Series
[DNLM: 1. Asthma–physiopathology, 2. Gene Expression Regulation. 3. Inflammation–
diagnosis. 4. Lung Diseases, Obstructive–pathophysiology. WF 553 H918 2001]
RC732.H85 2001
616.2'38075–dc21 00-058136
Preface
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. Vasodila-
tation underlies the flare and heat, plasma exudation the swelling, and activa-
tion of sensory nerves relays pain. In chronic conditions, skin biopsies show
inflammatory cell infiltrate. Inflammation is not unique to the skin and contrib-
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 un-
pleasant chest sensations associated with asthma attacks. Understanding the patho-
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.
Human Airway Inflammation: Sampling Techniques and Analytical
Protocols attempts to draw together many of the important methodologies
and protocols for assessing inflammation in human airways. We start with
techniques for the collection of samples. These can be such airway liquids
as sputum or exhaled gases as nitric oxide (NO). Once collected, the samples
can be used for the isolation and characterization of cells or for the measure-
ment of markers of inflammation. Techniques to isolate and analyze all of the
major inflammatory cells associated with lung inflammation are given. Similarly,
protocols to measure many of the inflammatory mediators and enzymes released
during lung inflammation are detailed herein. In an attempt to be inclusive and to
attract both experienced researchers and the novice, we have included specialized
chapters (for example, on tracing intracellular mediator storage), mixed with
v
vi Preface
more general chapters (for example, on Northern blotting). We wanted our vol-
ume to have something for everyone interested in assessing lung inflammation.
To achieve our aim, we are indebted to our authors for sharing their meth-
odological secrets. As we all know, following the published “recipe” does not
always (some would say ever) reproduce the data presented. To that end, the
“Methods” series includes the authors “Notes.” These are the tips and nuances
that convert abject failure into a successful experiment. Our contributors have
been most forthcoming on their methods, most of which were painstakingly
developed over long periods of time. Another factor that increases the impact
of the book is the inclusion of color figures in some of the chapters. We are
most grateful to Astra Zeneca and Boehringer Ingelheim, UK for sponsoring
color reproduction. Finally, we wish to thank John Walker and Thomas Lanigan
for inviting us to put this volume together—it has been an invaluable learning
experience for us all.
Duncan F. Rogers
Louise E. Donnelly
Contents
Preface ............................................................................................................v
Contributors....................................................................................................xi
1 Airway Inflammation and Remodeling in Asthma:
Current Concepts
Stephen T. Holgate................................................................................1
PART I: SAMPLE COLLECTION:INVASIVE TECHNIQUES
2 Biopsy Techniques: Optimization for Collection and Preservation
Marina Saetta and Graziella Turato...................................................19
3 Bronchoalveolar Lavage (BAL): Critical Evaluation of Techniques
Christopher Ward and E. Haydn Walters..........................................31
4 Nasal Secretions and Exudations: Collection and Approaches
to Analysis
Lennard Greiff, Morgan Andersson, and Carl G.A. Persson.........61
PART II. SAMPLE COLLECTION: NONINVASIVE TECHNIQUES
5 Induced Sputum: Selection Method
Margaret M. Kelly, Ann Efthimiadis, and
Frederick E. Hargreave...................................................................77
6 Induced Sputum: Whole Sample
Vera M. Keatings and Julia A. Nightingale.......................................93
7 Measurement of Exhaled Nitric Oxide and Carbon Monoxide
Sergei A. Kharitonov...........................................................................99
8 Measurement of Exhaled Hydrocarbons
Paolo Paredi, Sergei A. Kharitonov, and Peter J. Barnes............109
9 Breath Condensate as a Vehicle for Collection of Inflammatory
Mediators, Especially Hydrogen Peroxide
Johan C. de Jongste, Rijn Jöbsis, and H. Rolien Raatgeep........119
PART III. CELL ISOLATIONAND CULTURE
10 Airway Epithelial Cells (Primaries vs Cell Lines)
Louise E. Donnelly.............................................................................127
11 Isolation and Culture of Human Alveolar Type II Pneumocytes
Ian R. Witherden and Teresa D. Tetley...........................................137
vii
viii Contents
12 Pulmonary Artery Endothelial Cells
Joachim Seybold and Norbert Suttorp...........................................147
13 Isolation and Culture of Human Airway Smooth Muscle Cells
Aili L. Lazaar and Reynold A. Panettieri, Jr...................................155
14 Isolation and Purification of Human Mast Cells and Basophils
Bernhard F. Gibbs and Madeleine Ennis .......................................161
15 Neutrophils: Collection, Separation, and Activation
Sarah V. Culpitt..................................................................................177
16 Macrophages: Identification, Separation, and Function
Leonard W. Poulter and C.M. Burke................................................191
17 Collection, Separation, and Activation of Human T Lymphocytes
Yannis Sotsios and Stephen G. Ward.............................................205
18 Eosinophils: Collection, Separation, and Activation
Leo Koenderman, Jan van der Linden, Laurien Ulfman,
and Paul Coffer..............................................................................217
PART IV. GENE EXPRESSION/REGULATION
19 Analysis of Gene Expression: Reverse Transcription-Polymerase
Chain Reaction
El-Bdaoui Haddad and Jonathan Roussell....................................229
20 Analysis of Gene Expression: Northern Blotting
Jonathan Roussell and El-Bdaoui Haddad....................................247
21 The Technique of In Situ Hybridization
Sun Ying and A. Barry Kay..............................................................263
22 Measurement of Airway Mucin Gene Expression
Kelly Pritchard, Alinka K. Smith, and Duncan F. Rogers............285
23 Transcription Factors and Inflammatory Gene Regulation: Strategic
Approaches
Robert Newton and Karl J. Staples............................................... 295
PART V. MEDIATORS:EXPRESSIONAND FUNCTION
24 Inflammatory Mediators in Spontaneously Produced Sputum
Adam Hill, Simon Gompertz, Darren Bayley,
and Robert Stockley.....................................................................317
25 Flow Cytometry: Measurement of Leukocyte Receptor
Expression and Function
Adele Hartnell....................................................................................335
Contents ix
26 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.....................................................................345
27 Current In Vitro Models of Leukocyte Migration: Methods and
Interpretation
Jennifer R. Allport, Guillermo Garcia-Cardeña, Yaw-Chyn Lim,
and Francis W. Luscinskas.........................................................357
28 Tracing Intracellular Mediator Storage and Mobilization
in Eosinophils
Salahaddin Mahmudi-Azer, Paige Lacey,
and Redwan Moqbel.....................................................................367
29 Measurement of Metalloproteinases
Steven D. Shapiro, Diane Kelley and Dale Kobayashi.................383
30 Immunocytochemistry and Immunohistochemistry
Jochen Springer and Axel Fischer.................................................391
31 The Measurement of Cysteinyl Leukotrienes in Urine
Jay Y. Wescott and Sally E. Wenzel...............................................399
32 Quantitative Analysis of Cyclooxygenase Metabolites
of Arachidonic Acid
Ryszard Dworski, James R. Sheller,
and Brian W. Christman...............................................................411
33 Quantitative Analysis of F Isoprostanes
2
Ryszard Dworski, Brian W. Christman,
and James R. Sheller...................................................................423
Index............................................................................................................431
Contributors
JENNIFER R. ALLPORT • Vascular Research Division, Department of Pathology,
Brigham and Women’s Hospital, Boston, MA
MORGAN ANDERSSON • Department of Otorhinolaryngology, Head and Neck
Surgery, University Hospital, Lund, Sweden
PETER J. BARNES • Department of Thoracic Medicine, National Heart and
Lung Institute, Imperial College School of Science, Technology and
Medicine, London, UK
DARREN BAYLEY • Department of Respiratory Medicine, Queen Elizabeth
Hospital, Birmingham, UK
SHANNON A. BRYAN • Royal Brompton Clinical Studies Unit, National Heart
and Lung Institute, Imperial College School of Science, Technology and
Medicine, London, UK
C. M. BURKE • Department of Respiratory Medicine, James Connolly
Memorial Hospital, Blanchardstown, Dublin, Ireland
BRIAN W. CHRISTMAN • Center for Lung Research, Vanderbilt University
Medical Center, Nashville, TN
PAUL COFFER • Department of Pulmonary Diseases, University Hospital,
Utrecht, The Netherlands
SARAH V. CULPITT • Department of Thoracic Medicine, National Heart and
Lung Institute, Imperial College School of Science, Technology and
Medicine, London, UK
LOUISE E. DONNELLY • Department of Thoracic Medicine, National Heart
and Lung Institute, Imperial College School of Science, Technology and
Medicine, London, UK
RYSZARD DWORSKI • Division of Allergy, Pulmonary, and Critical Care
Medicine, Vanderbilt University School of Medicine, Nashville, TN
ANN EFTHIMIADIS • Firestone Regional Chest and Allergy Unit, St. Joseph’s
Hospital, Ontario, Canada
MADELEINE ENNIS • Department of Clinical Biochemistry, Institute of Clinical
Science, The Queen’s University of Belfast, Belfast, UK
AXEL FISCHER • Allergy Research Group, Humboldt University, Charité,
Berlin, Germany
GUILLERMO GARCIA-CARDEÑA • Vascular Research Division, Department of
Pathology, Brigham and Women’s Hospital, Boston, MA
xi