Superantigen Protocols

259 Pages·2003·1.91 MB·English

by  Matthew D. Baker

Superantigen Protocols
M E T H O D S I N M O L E C U L A R B I O L O G YTM
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M E T H O D S I N M O L E C U L A R B I O L O G YTM
Superantigen
Protocols
Edited by
Teresa Krakauer
Uniformed Services University of the Health Sciences, Bethesda, MD
US Army Medical Research Institute of Infectious Diseases,
Frederick, MD
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Cover illustration: From Fig. 3B in Chapter 5 by S. Munir Alam and Nicholas R. J. Gascoigne; from Fig. 2
in Chapter 11 by Teresa Krakauer, Xin Chen, O. M. Zack Howard, and Howard A. Young; and from Fig. 2
in Chapter 12 by Lars Björk.
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Main entry under title: Methods in molecular biology™.
Superantigen protocols/edited by Teresa Krakauer
p.;cm.--(Methods in molecular biology; 214)
Includes bibliographical references and index.
ISBN 0-89603-984-6 (alk. paper)
1. Superantigens--Laboratory manuals.
[DNLM: 1. Superantigens. 2. Clinical Laboratory Techniques. QW 573
S958 2002] I. Krakauer, Teresa. II. Methods in molecular biology
(Clifton, N.J.); v. 214.
QR186.6.S94 S867 2002
616.07'92--dc21 2002017291
Dedication
To my super-family for providing me with an enriched environment to
study and learn.
Teresa Krakauer, PhD
v
Preface
Superantigen Protocols assembles experimental protocols that have proved
useful for the study of superantigens. These techniques will allow researchers
from various areas of cell biology, microbiology, immunology, biochemistry,
and molecular biology to assess the physical characteristics and biological
effects of well-known superantigens as well as of putative substances that
might have superantigenic activities, and to explore therapies for superantigen-
induced effects.
Microbial exotoxins have been studied for decades as virulence factors
because of their pathogenic effects. The term “superantigen” was coined by
Marrack and Kappler a decade ago for some of these molecules because of
their potent T-cell stimulatory activities. In recent years, advances in molecu-
lar biology provide recombinant as well as natural superantigens in highly
purified form for physical characterization. Superantigens are now used
extensively as tools to study interactions between receptors on cells of the
immune system as they bind to major histocompatibility complex class II mol-
ecules on antigen-presenting cells and V regions of T-cell receptors. The bio-
β
logical effects that result from these interactions are studied both in vitro and
in vivo. The intent of this book is, therefore, to bring together up-to-date tech-
niques developed by experts in the field of biochemistry, immunology, and
molecular biology for the study of superantigens.
Superantigen Protocols begins with an overview of the field to provide
background information on the various classes of superantigens and their struc-
ture. This summary is followed by a group of chapters on methods to define
binding and other physical interactions of superantigens with their cellular
receptors. Finally, techniques used in the study of the biological effects of
superantigens are covered to guide researchers in the assessment of cellular
activities and molecules induced by superantigens. In addition, approaches to
the study and the development of therapeutics in suppressing the superantigen-
induced effects are presented.
Teresa Krakauer, PhD
vii
Contents
Dedication.........................................................................................................v
Preface ...........................................................................................................vii
Contributors..................................................................................................xi
1 Superantigens: Structure, Function, and Diversity
Matthew D. Baker and K. Ravi Acharya..............................................1
2 Expression, Purification, and Detection of Novel Streptococcal
Superantigens
John K. McCormick and Patrick M. Schlievert................................33
3 Flow Cytometric Detection of MMTV Superantigens
Gary Winslow.......................................................................................45
4 Spectrophotometric Methods for the Determination
of Superantigen Structure and Stability
Anders Cavallin, Karin Petersson, and Göran Forsberg................55
5 Binding Kinetics of Superantigen with TCR and MHC Class II
S. Munir Alam and Nicholas R. J. Gascoigne..................................65
6 Directed Evolution of T-Cell Receptors for Binding Superantigens
Hywyn R. O. Churchill and David M. Kranz......................................87
7 Analysis of Superantigen Binding to Soluble T-Cell Receptors
Hywyn R. O. Churchill and David M. Kranz....................................101
8 Role of Accessory Molecules in the Superantigen-Induced
Activation of Peripheral Blood T Cells
Catherine Gelin, Marie-Thérèse Zilber,
and Dominique Charron...............................................................113
9 T-Lymphocyte Activation Induced by Staphylococcal
Enterotoxin Superantigens: Analysis of Protein Tyrosine
Phosphorylation
Anne Roumier, Florence Niedergang, and Andrés Alcover.........127
10 Measurement of Proinflammatory Cytokines and T-Cell
Proliferative Response in Superantigen-Activated Human
Peripheral Blood Mononuclear Cells
Teresa Krakauer.................................................................................137
ix
x Contents
11 RNase Protection Assay for the Study of the Differential Effects
of Therapeutic Agents in Suppressing Staphylococcal
Enterotoxin B-Induced Cytokines in Human Peripheral
Blood Mononuclear Cells
Teresa Krakauer, Xin Chen, O. M. Zack Howard,
and Howard A. Young...................................................................151
12 Staining Protocol for Superantigen-Induced Cytokine Production
Studied at the Single-Cell Level
Lars Björk...........................................................................................165
13 Assessment of Specific T-Cell Activation by Superantigens
Natalie Sutkowski and Brigitte T. Huber........................................185
14 Superantigen-Induced Changes in Epithelial Ion Transport
and Barrier Function: Use of an In Vitro Model
James L. Watson and Derek M. McKay...........................................219
15 Pyrogenic, Lethal, and Emetic Properties of Superantigens
in Rabbits and Primates
John K. McCormick, Gregory A. Bohach,
and Patrick M. Schlievert.............................................................245
Index............................................................................................................255
Contributors
K. RAVI ACHARYA• Department of Biology and Biochemistry, University
of Bath, Claverton Down, Bath, UK
S. MUNIR ALAM• Division of Rheumatology, Allergy and Clinical Immunology,
Department of Medicine, Duke University Medical Center, Durham, NC
ANDRÉS ALCOVER• Unité de Biologie des Interactions Cellulaires, Institut
Pasteur, Paris, France
MATTHEW D. BAKER• Department of Biology and Biochemistry, University
of Bath, Claverton Down, Bath, UK
LARS BJÖRK• Department of Biopharmaceutical Development, Biovitrum,
Stockholm, Sweden
GREGORY A. BOHACH• Department of Microbiology, Molecular Biology
and Biochemistry, University of Idaho, Moscow, ID
ANDERS CAVALLIN• Department of Molecular Biology, Astra Zeneca,
Mölndel, Sweden
DOMINIQUE CHARRON• Inserm U396, Institut Biomedical des Cordeliers
Université, Paris, France
XIN CHEN• Laboratory of Molecular Immunoregulation, National Cancer
Institute, Frederick, MD
HYWYN R. O. CHURCHILL• Department of Biochemistry, University of Illinois,
Urbana, IL
GÖRAN FORSBERG• Department of Biology, Active Biotech Research, Lund,
Sweden
NICHOLAS R. J. GASCOIGNE• Department of Immunology, The Scripps Research
Institute, La Jolla, CA
CATHERINE GELIN• Inserm U396, Inst d’Hématologie, Hôpital Saint-Louis,
Paris, France
O. M. ZACK HOWARD• Laboratory of Molecular Immunoregulation,
National Cancer Institute, Frederick, MD
BRIGITTE T. HUBER• Department of Pathology, Tufts University School
of Medicine, Boston, MA
TERESA KRAKAUER• Department of Preventive Medicine and Biometrics,
Uniformed Services University of the Health Sciences, Bethesda, MD;
Department of Immunology and Molecular Biology, United States Army
Medical Research Institute of Infectious Diseases, Fort Detrick,
Frederick, MD
xi