Table Of ContentM 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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Adoptive Immunotherapy
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
111133.Multiple Myeloma: Methods and Protocols, 9966. Hepatitis B and D Protocols: Volume 2,
edited by Ross D. Brown and P. Joy Ho, Immunology, Model Systems, and Clinical
2005 Studies, edited by Robert K. Hamatake and
Johnson Y. N. Lau, 2004
111122.Molecular Cardiology: Methods and
Protocols, edited by Zhongjie Sun, 2005 9955. Hepatitis B and D Protocols: Volume 1,
Detection, Genotypes, and
111111.Chemosensitivity: Volume 2, In Vivo
Characterization, edited by Robert K.
Models, Imaging, and Molecular Regulators,
Hamatake and Johnson Y. N. Lau, 2004
edited by Rosalyn D. Blumethal, 2005
9944. Molecular Diagnosis of Infectious
111100.Chemosensitivity: Volume 1, In Vitro Diseases, Second Edition, edited by Jochen
Assays, edited by Rosalyn D. Blumethal, Decker and Udo Reischl, 2004
2005
9933. Anticoagulants, Antiplatelets, and
110099.Adoptive Immunotherapy, Methods and Thrombolytics, edited by Shaker A.
Protocols, edited by Burkhard Ludewig and Mousa, 2004
Matthias W. Hoffmann, 2005
9922. Molecular Diagnosis of Genetic Diseases,
110088.Hypertension, Methods and Protocols, Second Edition, edited by Rob Elles and
edited by Jérôme P. Fennell and Andrew H. Roger Mountford, 2004
Baker, 2005 9911. Pediatric Hematology: Methods and
110077.Human Cell Culture Protocols, Second Protocols, edited by Nicholas J. Goulden
Edition, edited by Joanna Picot, 2005 and Colin G. Steward, 2003
110066.Antisense Therapeutics, Second Edition, 9900. Suicide Gene Therapy: Methods and
edited by M. Ian Phillips, 2005 Reviews, edited by Caroline J. Springer, 2004
110055.Developmental Hematopoiesis: Methods 8899. The Blood–Brain Barrier: Biology and
and Protocols, edited by Margaret H. Research Protocols, edited by Sukriti Nag,
Baron, 2005 2003
110044.Stroke Genomics: Methods and Reviews, 8888. Cancer Cell Culture: Methods and
edited by Simon J. Read and David Virley, Protocols, edited by Simon P. Langdon, 2003
2005 8877. Vaccine Protocols, Second Edition, edited
110033.Pancreatic Cancer: Methods and Protocols, by Andrew Robinson, Michael J. Hudson,
edited by Gloria H. Su, 2005 and Martin P. Cranage, 2003
110022.Autoimmunity: Methods and Protocols, 8866. Renal Disease: Techniques and Protocols,
edited by Andras Perl, 2004 edited by Michael S. Goligorsky, 2003
110011.Cartilage and Osteoarthritis: Volume 2, 8855. Novel Anticancer Drug Protocols, edited
Structure and In Vivo Analysis, edited by by John K. Buolamwini and Alex A. Adjei,
Frederic De Ceuninck, Massimo Sabatini, 2003
and Philippe Pastoureau, 2004 8844. Opioid Research: Methods and Protocols,
110000.Cartilage and Osteoarthritis: Volume 1, edited by Zhizhong Z. Pan, 2003
Cellular and Molecular Tools, edited by 8833. Diabetes Mellitus: Methods and Protocols,
Massimo Sabatini, Philippe Pastoureau, edited by Sabire Özcan, 2003
and Frederic De Ceuninck, 2004
8822. Hemoglobin Disorders: Molecular
9999. Pain Research: Methods and Protocols, Methods and Protocols, edited by Ronald
edited by David Z. Luo, 2004 L. Nagel, 2003
9988. Tumor Necrosis Factor: Methods and 8811. Prostate Cancer Methods and Protocols,
Protocols, edited by Angelo Corti and edited by Pamela J. Russell, Paul Jackson,
Pietro Ghezzi, 2004 and Elizabeth A. Kingsley, 2003
9977. Molecular Diagnosis of Cancer: Methods 8800. Bone Research Protocols, edited by Miep
and Protocols, Second Edition, edited by H. Helfrich and Stuart H. Ralston, 2003
Joseph E. Roulston and John M. S. Bartlett, 7799. Drugs of Abuse: Neurological Reviews and
2004 Protocols, edited by John Q. Wang, 2003
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 E™
Adoptive
Immunotherapy
Methods and Protocols
Edited by
Burkhard Ludewig
Research Department, Kantonsspital St. Gallen, Switzerland
and
Matthias W. Hoffmann
Department of Abdominal and Transplant Surgery,
Hannover Medical School, Hannover, Germany
© 2005 Humana Press Inc.
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www.humanapress.com
All rights reserved.
No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any
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Production Editor: Tracy Catanese
Cover design by Patricia F. Cleary
Cover illustration: Figure 1 from Chapter 3, “Delivery of Tumor Antigens to Dendritic Cells Using Biode-
gradable Microspheres,” by Ying Waeckerle-Men, Bruno Gander, and Marcus Groettrup. Schematic
representation of the regulation of dendritic cell persistence within secondary lymphoid organs by effector
T-cells. Immune responses are positively regulated by dendritic cells mediating antigen influx into secondary
lymphoid organs from peripheral sites such as tumor tissues.
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Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1
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Library of Congress Cataloging-in-Publication Data
Adoptive immunotherapy : methods and protocols / edited by Burkhard Ludewig and Matthias W. Hoffmann.
p. ; cm. -- (Methods in molecular medicine ; 109)
Includes bibliographical references and index.
ISBN 1-58829-406-4 (alk. paper)
1. Cancer--Immunotherapy. 2. Cellular therapy. 3. Immunocompetent cells--Therapeutic use.
[DNLM: 1. Immunotherapy, Adoptive--methods. 2. Antibodies--therapeutic use. 3. Cytotoxicity,
Immunologic. 4. Dendritic Cells. 5. Stem Cell Transplantation. 6. T-Lymphocytes.
QW 940 A239 2005] I. Ludewig, Burkhard. II. Hoffmann, Matthias W. III. Series.
RC271.I45A335 2005
616.99'4061--dc22
2004010540
Preface
Over the last decade, the advances in cellular and molecular immunology have
been tremendous. Our continuously improving understanding of the immune
system and the appreciation of the mechanisms by which tumors and viral or
bacterial infections are controlled have led to promising new treatment strate-
gies. Adoptive transfer of tailored antigen-specific immune cells and/or opti-
mally designed immunological effector molecules is an elegant and promising
approach to the establishment or restoration of protective immune responses. At
this point, it appears timely to publish the present volume on Adoptive Immuno-
therapy for the Methods in Molecular Medicine™ series.
Experts from various fields contributed to this comprehensive collection of
state-of-the-art methods for adoptive immunotherapy. Recent technical advances
and specific methods from the areas of dendritic cell therapy, generation of anti-
gen-specific cytotoxic T cells, and antitumor treatment with specific antibodies
have been compiled. Particular emphasis is placed on preclinical and clinical
applications. These chapters are complemented by articles describing the
progress of selected approaches in clinical trials. Leading experts in the field
provide theoretical overviews and point out future directions for the improve-
ment of adoptive immunotherapy. Additional chapters on the molecular defini-
tion of target antigens, mathematical modeling approaches to immunotherapy,
and the utilization of regulatory T cells complete the volume. We were fortunate
to receive support from leading scientists from all over the world who contrib-
uted their most recent experimental protocols and shared their practical advice.
Special emphasis was placed on the “Notes” section to include helpful trouble-
shooting advice for daily work at the lab bench, which is a unique feature of the
Methods in Molecular Medicine™ series. It is our hope that we have achieved
our main objective to provide helpful information for scientists just entering the
field of adoptive immunotherapy. Furthermore, this volume attempts to encour-
age more experienced researchers to try novel experimental approaches.
We are indebted to all our authors for generously making their time avail-
able in contributing to this volume and for the excellent quality of their work.
Marcel Kremer’s help in the final editing process is greatly appreciated.
Finally, we want to thank the Series Editor, John Walker, and the staff of
Humana Press for their support and continuous encouragement.
Burkhard Ludewig
Matthias Hoffmann
v
vi Preface
Contents
Preface ..............................................................................................................v
Contributors .....................................................................................................xi
1 Exploiting Dendritic Cells for Active Immunotherapy
of Cancer and Chronic Infection
David O’Neill and Nina Bhardwaj.......................................................1
2 A Mathematical Approach for Optimizing Dendritic
Cell-Based Immunotherapy
Gennady Bocharov, Neville J. Ford, and Burkhard Ludewig.............19
3 Delivery of Tumor Antigens to Dendritic Cells
Using Biodegradable Microspheres
Ying Waeckerle-Men, Bruno Gander, and Marcus Groettrup...........35
4 RNA Transfection of Dendritic Cells
Frank Grünebach, Martin R. Müller, and Peter Brossart...................47
5 Isolation and Generation of Clinical-Grade Dendritic Cells
Using the CliniMACS System
John D. M. Campbell, Christoph Piechaczek, Gregor Winkels,
Edith Schwamborn, Daniela Micheli, Sonja Hennemann,
and Jürgen Schmitz.........................................................................55
6 Generation of Clinical Grade Monocyte-Derived Dendritic Cells
Using the CliniMACS System
Thomas Putz, Hubert Gander, Reinhold Ramoner,
Claudia Zelle-Rieser, Andrea Rahm, Walter Nussbaumer,
Georg Bartsch, Lorenz Höltl, and Martin Thurnher......................71
7 Adenoviral Transduction of Dendritic Cells
Rienk Offringa, Kitty Kwappenberg, Martijn Rabelink,
Delphine Rea, and Rob Hoeben.....................................................83
8 Generation of Autologous Peptide- and Protein-Pulsed
Dendritic Cells for Patient-Specific Immunotherapy
David O’Neill and Nina Bhardwaj.....................................................97
9 Phenotypical and Functional Characterization
of Clinical-Grade Dendritic Cells
I. Jolanda M. de Vries, Gosse J. Adema, Cornelis J. A. Punt,
and Carl G. Figdor........................................................................113
10 Dendritic Cells in Clinical Trials for Multiple Myeloma
Volker L. Reichardt and Peter Brossart............................................127
vii
viii Contents
11 Identification of Tumor-Associated Autoantigens With SEREX
Özlem Türeci, Dirk Usener, Sandra Schneider,
and Ugur Sahin.............................................................................137
12 Application of Proteomics and Protein Analysis for Biomarker
and Target Finding for Immunotherapy
Petra Weingarten, Petra Lutter, Andreas Wattenberg,
Martin Blueggel, Sonja Bailey, Joachim Klose,
Helmut E. Meyer, and Christoph Huels.......................................155
13 Isolation and Expansion of Tumor-Reactive Cytotoxic
T-Cell Clones for Adoptive Immunotherapy
Helga Bernhard, Burkhard Schmidt, Dirk H. Busch,
and Christian Peschel...................................................................175
14 Tracking Adoptively Transferred Antigen-Specific T-Cells
With Peptide/MHC Multimers
Norbert Meidenbauer and Andreas Mackensen...............................185
15 Gene Transfer of MHC-Restricted Receptors
Helmut W. H. G. Kessels, Monika C. Wolkers,
and Ton N. M. Schumacher..........................................................201
16 Immunotherapy With CTL Restricted by Nonself MHC
Liquan Gao, Anne-Marie Downs, and Hans J. Stauss.......................215
17 Designing TCR for Cancer Immunotherapy
Ralf-Holger Voss, Jürgen Kuball, Matthias Theobald.......................229
18 Isolation and Expansion of Tumor-Specific CD4+ T-Cells
by Means of Cytokine Secretion
Christian Becker................................................................................257
19 Methods for the Ex Vivo Characterization of Human CD8+ T
Subsets Based on Gene Expression and Replicative
History Analysis
Nathalie Rufer, Patrick Reichenbach, and Pedro Romero...............265
20 Regulatory T Cells in Antitumor Therapy: Isolation
and Functional Testing of CD4+CD25+ Regulatory T-Cells
Helmut Jonuleit and Edgar Schmitt..................................................285
21 Monoclonal Antibody-Based Strategies in Autoimmunity
and Transplantation
Lucienne Chatenoud.........................................................................297
22 Producing Bispecific and Bifunctional Antibodies
Dipankar Das and Mavanur R. Suresh..............................................329
23 Antibody–Cytokine Fusion Proteins for the Therapy of Cancer
Gustavo Helguera and Manuel L. Penichet......................................347
Contents ix
24 Synthesis and Biological Evaluation of a Paclitaxel
Immunoconjugate
Ahmad Safavy and Kevin P. Raisch...................................................375
25 Cytotoxic Tumor Targeting With scFv Antibody-Modified
Liposomes
Cornelia Marty and Reto A. Schwendener.......................................389
26 Intravenous Immunoglobin Treatment for Fibrosis,
Atherosclerosis, and Malignant Conditions
Ilan Krause and Yehuda Shoenfeld...................................................403
27 Study of T-Cell Costimulatory Blockade In Vivo
at a Single Cell Level
Minh Diem Vu and Xian Chang Li....................................................409
28 Stem Cell Transplantation: Graft-Mediated Antileukemia Effects
William J. Hogan and Hans Joachim Deeg.......................................421
29 Influence of Radiation Protocols on Graft-vs-Host Disease
Incidence After Bone-Marrow Transplantation
in Experimental Models
Sebastian Schwarte and Matthias W. Hoffmann..............................445
30 Induction of Chimerism and Tolerance Using Freshly Purified
or Cultured Hematopoietic Stem Cells
in Nonmyeloablated Mice
Nikos Emmanouilidis and Christian P. Larsen..................................459
31 Induction of Mixed vs Full Chimerism to Potentiate GVL
Effects After Bone-Marrow Transplantation
Markus Y. Mapara and Megan Sykes................................................469
32 Application of Donor Lymphocytes Expressing a Suicide Gene
for Early GVL Induction and Later Control of GVH Reactions
After Bone-Marrow Transplantation
Attilio Bondanza, Fabio Ciceri, and Chiara Bonini..........................475
Index............................................................................................................487
