Septic Shock Methods and Protocols

205 Pages·2000·0.938 MB·English

by  Paul A. Ketchum, Thomas J. Novitsky (auth.), Thomas J. Evans (eds.)

Septic Shock Methods and Protocols
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
John M. Walker, SERIES EDITOR
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M E T H O D S I N M O L E C U L A R M ED I C I N ETM
Septic Shock
Methods and
Protocols
Edited by
Thomas J. Evans
Imperial College School of Medicine
London, UK
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Library of Congress Cataloging-in-Publication Data
Septic shock methods and protocols / edited by Thomas J. Evans.
p. cm. -- (Methods in molecular medicine ; 36)
Includes bibliographical references and index.
ISBN 0-89603-730-4 (alk. paper)
1. Septic shock--Research--Methodology. 2. Endotoxins--Research--Methodol-
ogy. 3. Cytokines--Research--Methodology. I. Evans, Thomas J., 1959. II. Series.
[DNLM: 1. Shock, Septic--immunology. 2. Cell Culture--methods.
3. Cytokines--immunology. 4. Endotoxins--isolation & purification.
5. Nitric Oxide--antagonists & inhibitors. QZ 140 S4796 2000]
RC182.S4S466 2000
6169.047--dc21
DNLM/DLC
for Library of Congress 99-29663
CIP
Preface
Septic shock remains a serious medical condition with high mortality.
Despite many advances in intensive care medicine and antibiotic develop-
ment, this has not changed appreciably in the last 20 years. Frustratingly, over
the same period of time, enormous advances have been made in understand-
ing the underlying pathogenic mechanisms of this condition. This has resulted
in the development of several novel therapies for septic shock, which, despite
excellent theoretical grounds for their efficacy, have failed in altering mortal-
ity attributable to sepsis.
The reasons for these failures are multiple, but it is clear that further
research is required aimed at increasing our understanding of the basic patho-
physiological processes that occur following infection. Research into septic
shock draws upon a number of different disciplines, ranging from molecular
and cellular biology to physiological measurements on whole animals. Septic
Shock Methods and Protocolsis an attempt to draw together into one volume
a number of protocols that are of use in the investigation of the mechanisms of
septic shock. I have divided the book into five sections. The first deals with
endotoxin, the lipopolysaccharide component of the Gram-negative cell mem-
brane that can mimic many of the features of septic shock. Gram-positive
organisms are found increasingly as causes of septic shock, and several as-
pects of toxins produced from these bacteria are considered in the second sec-
tion. Cytokines have been a central focus of interest in sepsis research for
many years and several aspects of cytokine biology are highlighted in the third
section. In the fourth section, methods for studying nitric oxide and other re-
active nitrogen intermediates are considered. Finally, the last section describes
a variety of methods for studying primary cell cultures, an essential compo-
nent of developing in vitro methods to study septic shock.
Obviously Septic Shock Methods and Protocols cannot provide an ex-
haustive account of every protocol that might be used in sepsis research. I
have therefore carefully chosen those highlights that I believe either are of
great intrinsic importance or are poorly covered elsewhere in published proto-
col manuals. I am very grateful to all the authors who have contributed to this
v
vi Preface
volume. I believe their collected experience is invaluable and I hope that this
book will allow both newcomers and those with more experience to apply
successfully the techniques needed for their research.
T. J. Evans
Contents
Preface ............................................................................................................v
Contributors....................................................................................................ix
PART 1 ENDOTOXIN
1 Assay of Endotoxin by Limulus Amebocyte Lysate
Paul A. Ketchum and Thomas J. Novitsky.........................................3
2 Preparation of Endotoxin from Pathogenic Gram-Negative Bacteria
Alexander Shnyra, Michael Luchi, and David C. Morrison............13
3 Assay of Anti-Endotoxin Antibodies
Lore Brade............................................................................................27
4 Isolation of the Bactericidal/Permeability-Increasing Protein
from Polymorphonuclear Leukocytes
by Reversible Binding to Target Bacteria
Jerrold Weiss.......................................................................................37
5 Purification of Lipopolysaccharide Binding Protein
Didier Heumann...................................................................................45
PART 2 OTHER BACTERIAL PRODUCTS
6 Purification of Streptococcal Pyrogenic Exotoxin A
Manuela Roggiani and Patrick M. Schlievert...................................59
7 Assay for Superantigens
Shiranee Sriskandan...........................................................................67
PART 3 CYTOKINES
8 Bioassay for Tumor Necrosis Factors-α and β
Thomas J. Evans.................................................................................83
9 Assay of Soluble Tumor Necrosis Factor Receptors
Maarten G. Bouma and Wim A. Buurman.........................................91
10 Whole-Blood Assays for Cytokine Production
Daniel G. Remick, David E. Newcomb, and Jon S. Friedland......101
vii
viii Contents
PART 4 NITRIC OXIDE AND OTHER REACTIVE NITROGEN INTERMEDIATES
11 Nitric Oxide Synthase Inhibitors
Dilani K. Siriwardena, Hajime Tagori,
Christoph Thiemermann...............................................................115
12 In Situ Detection of Nitric Oxide
Tadeusz Malinski...............................................................................133
13 Immunochemical Detection of Nitric Oxide Synthase
in Human Tissue
Lee D. K. Buttery and Julia M. Polak...............................................145
14 Anti-Nitrotyrosine Antibodies for Immunohistochemistry
Liliana Viera, Yao Zu Ye, and Joseph S. Beckman.......................159
15 Detection of Peroxynitrite in Biological Fluids
Stuart Malcolm, Raymond Foust III, Caryn Hertkorn,
and Harry Ischiropoulos...............................................................171
PART 5 CELL CULTURE TECHNIQUES
16 Myocardial Cells in Culture to Study Effects of Cytokines
Kevin A. Krown and Roger A. Sabbadini........................................181
17 Culture of Primary Human Bronchial Epithelial Cells
Joanna Picot.......................................................................................189
18 Primary Culture of Human Proximal Renal Tubular Epithelial Cells
Paul A. Glynne....................................................................................197
Index............................................................................................................207
C
ONTRIBUTORS
JOSEPH S. BECKMAN • Departments of Anesthesiology, Biochemistry and
Molecular Genetics, and Neurobiology and The UAB Center for Free
Radical Biology, The University of Alabama at Birmingham,
Birmingham, AL
MAARTEN G. BOUMA • Department of Surgery, Maastricht University,
Maastricht, The Netherlands
LEE D. K. BUTTERY • Department of Histochemistry, Imperial College
School of Medicine, London, UK
LORE BRADE • Research Center Borstel, Center for Medicine
and Biosciences, Borstel, Germany
WIM A. BUURMAN • Department of Surgery, Maastricht University,
Maastricht, The Netherlands
RAYMOND FOUST III • Stokes Research Institute and Department
of Biochemistry and Biophysics, Children’s Hospital of Philadelphia
and The University of Pennsylvania, Philadelphia, PA
THOMAS J. EVANS • Department of Infectious Diseases, Imperial College
School of Medicine, Hammersmith Hospital, London, UK
JON S. FRIEDLAND • Department of Infectious Diseases, Imperial College
School of Medicine, Hammersmith Hospital, London, UK
PAUL A. GLYNNE • Department of Infectious Diseases, Imperial College
School of Medicine, Hammersmith Hospital, UK
CARYN HERTKORN • Stokes Research Institute and Department
of Biochemistry and Biophysics, Children’s Hospital of Philadelphia
and The University of Pennsylvania, Philadelphia, PA
DIDIER HEUMANN • Division of Infectious Diseases, CHUV-1011, Lausanne,
Switzerland
HARRY ISCHIROPOULOS • Stokes Research Institute and Department
of Biochemistry and Biophysics, Children’s Hospital of Philadelphia
and The University of Pennsylvania, Philadelphia, PA
PAUL A. KETCHUM • Associates of Cape Cod, Falmouth, MA
KEVIN A. KROWN • Rees-Stealy Research Foundation, San Diego, CA
MICHAEL LUCHI • Department of Medicine, Division of Infectious Disease,
Kansas University Medical Center, Kansas City, KS
STUART MALCOLM • Stokes Research Institute and Department
of Biochemistry and Biophysics, Children’s Hospital of Philadelphia
and The University of Pennsylvania, Philadelphia, PA
ix
x Contributors
TADEUSZ MALINSKI• Center for Biomedical Research, Oakland University,
Rochester, Ml
DAVID C. MORRISON • Saint Luke’s Hospital/Shawnee Mission Health
System; Division of Basic Medical Science, School of Medicine,
University of Missouri-Kansas City, Kansas City, MO
DAVID E. NEWCOMB • Department of Pathology, University of Michigan,
Ann Arbor, MI
JOANNA PICOT • Department of Infectious Diseases, Imperial College School
of Medicine, Hammersmith Hospital, London, UK
THOMAS J. NOVITSKY • Associates of Cape Cod, Falmouth, MA
JULIA M. POLAK • Department of Histochemistry, Imperial College School
of Medicine, London, UK
DANIEL G. REMICK • Department of Pathology, University of Michigan,
Ann Arbor, MI
MANUELA ROGGIANI • Department of Medicine, Division of Infectious
Disease, Kansas University Medical Center, Kansas City, KS
ROGER A. SABBADINI • Department of Biology, San Diego State University,
San Diego, CA
PATRICK M. SCHLIEVERT • Department of Microbiology, University
of Minnesota, Minneapolis, MN
ALEXANDER SHNYRA • Department of Microbiology, Molecular Genetics
and Immunology, Kansas University Medical Center, Kansas City, KS
DILANI K. SIRIWARDENA • Moorfields Eye Hospital, London, UK
SHIRANEE SRISKANDAN • Department of Infectious Diseases, Imperial College
School of Medicine at Hammersmith Hospital, London, UK
HAJIME TAGORI • Department of Neonatology, Nagoya City University
Medical School, Nagoya, Japan
CHRISTOPH THIEMERMANN • The William Harvey Research Institute,
St. Bartholomew’s and the Royal London School of Medicine
and Dentistry, London, UK
LILIANA VIERA • Department of Anesthesiology and The UAB Center
for Free Radical Biology, The University of Alabama at Birmingham,
Birmingham, AL; Departamento de Histologia y Embriologia, Facultad
de Medicina, Universidad de la Republica, Montevideo, Uruguay
JERROLD WEISS • The Inflammation Program, Departments of Internal
Medicine and Microbiology, University of Iowa College of Medicine,
Iowa City, IA
YAO ZU YE • Department of Anesthesiology and The UAB Center
for Free Radical Biology, The University of Alabama at Birmingham,
Birmingham, AL