Intracellular Antibodies: Development and Applications

199 Pages·1997·5.62 MB·English

by  Antonino Cattaneo

Springer-Verlag Berlin Heidelberg GmbH
Antonino Cattaneo Silvia Biocca (Eds.)
Intracellular Antibodies:
Development and Applications
Springer
Antonino Cattaneo, Ph. D. Silvia Biocca, Ph.D.
Biophysics Sector Department of Experimental Medicine
International School for Advanced Studies and Biochemical Seiences
(SISSA) University of Rome "Tor Vergata"
Via Beirut 2/4 Via di Tor Vergata 135
34013 Trieste 00133 Roma
Italy Italy
ISBN 978-3-662-07994-2
Biotechnology Intelligence Unit
llbrary of Congress Cataloging-in-Publication Data
Cattaneo, Antonino, 1954-
Intracellular anubodies: developments and applications I Antonino Cattaneo, Silvia Biocca.
p.cm.- (Biotechnology intelligence unit)
Includes bibliographical references and index.
ISBN 978-3-662-o7994-2 ISBN 978-3-662-07992-8 (eBook)
DOI 10.1007/978-3-662-07992-8
1. Immunoglobulins. 2. Immunization. 3-Cellular immunity. 4-Immunogenetics.
I. Biocca, Silvia. 11. Series
[DNLM: 1. Immunity, Cellular-physiology. 2. Antibodies-physiology. 3· Antibody Formation-physiology.
4-Gene expression-physiology. QW 568.C3681 1997]
QR 186.7.C38 1997
616.o7'98-DC21 97-20286 CIP
DNLM/DLC
for Library of Congress
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sed tua me virtus tarnen et sperata voluptas
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Lucrezio Caro, De rerum natura (1,136-145)
PREFACE=====================
=====================
T
he concept of exploiting the virtually unlimited repertoire of anti
hodies by ectopically expressing antibody genes for functional stud
ies is ten years old. During this period, many developments have trans
formed this idea into a technologywith broad potential. Now the prom
ises, the problems and the perspectives can be more clearly identified.
This volume addresses these issues by describing what has been achieved
already, what could be conceivably achieved in the future and which
problems require solutions and improvements. The volume is intended
for people involved in different areas of research and therefore, where
possible, we have tried to describe the background to the technology
and to the applications.
The book reflects a personal view of how the field of ectopic anti
body expression has evolved and of the directions in which it may evolve.
Ideas are presented without constraints. The focus is on the general
concepts and ideas, but methodological aspects are covered as weil.
We are particularly grateful to our friend Roberto Sitia, whose
help, experience and lively discussions were vital for many aspects of
this work. Chapter 4 reflects in part his ideas, filtered through our own
eyes and biases.
We feel privileged to have received, for many years, the support
and guidance of Rita Levi-Montalcini, whose seminal experiments on
immunosympathectomy have provided a germ which has developed
into this work. We are very grateful to Rita for her support and
friendship.
We would like to express our deep gratitude to Pietro Calissano,
Director of the CNR Institute of Neurobiology (Roma), for his friendly
and magnanimous support throughout many years, in particular in the
initial phases of the development of this work.
A partiewar tribute goes to the researchers in our laboratories,
who have contributed to this rapidly moving field in the past few years
with their work, discussions, criticisms and high expectations, as weil
as to those who are starting now: Andrew Bradbury, Patrizia Piccioli,
Francesca Ruberti, Anna Di Luzio, Nicola Gargano, Stefania Gonfloni,
Luisa Fasulo, Michela Visentin, Massimo Righi, Lidija Persic, Marco
Tafani, Samantha Messina, Alessio Cardinale.
Special thanks to Paola Pierandrei-Amaldi, for her friendly and
enthusiastic scientific help, which established a turning point for us in
the development of the intraceilular antibody work.
The assistance of Samantha Messina and the cheerful collabora
tion ofVanessa Varnier in some phases of the preparation of this book
are gratefully acknowledged.
AC is very grateful to Cesar Milstein and Michael Neuherger for
thinking that the ectopic expression of antiborlies was worth pursuing,
and for allowing that pursuit to begin at the MRC Labaratory of Mo
lecular Biology ( Cambridge, UK). It was a pleasure to share Iab 310 with
Andrew and Michal. This book was partly written while AC was on
sabbaticalleave at the MRC Labaratory of Molecular Biology. AC wishes
to thank Terry Rabbitts and Greg Winter, Joint Heads of the PNAC Di
vision of the LMB, for the exciting opportunity.
SB thanks the Department of Experimental Medicine and Bio
chemical Seiences of the University of Rome "Tor Vergata" for assis
tance and support.
AC is grateful to Daniele Amati, Director of the International
School for Advanced Sturlies (SISSA), for his enthusiastic encourage
ment and support.
The research mentioned in the text, from the authors, laborato
ries, has been supported by research contracts with NE.FAC within the
National Research Plan Neurobiological Systems of the Ministero della
Ricerca Scientifica e Tecnologica, Consiglio Nazionale delle Ricerche
(CNR) (PF Biotecnologie,PF Ingegneria Genetica,PF Oncologia) INFM
and S.I.R.S. srl. Part of the work has been intermittently funded by the
Ministero della Sanita' (AIDS Project).
Finally, we would like to thank the staff of Landes Bioscience for
their help and understanding.
A. Cattaneo and S. Biocca
Roma, 3 April1997
r.:=====CONTENT~~====:=:::;-1
1. Intracellular and Intercellular Immunization .......................... 1
Antonino Cattaneo and Silvia Biocca
The Concept ofEctopic Antibody Expression for
Intracellular and Intercellular Immunization with
Antibody Genes ........................................................................ 1
Ectopic Antibody Expression: Different Cells and
Different Intracellular Compartments ................................... 2
Historical Overview ..................................................................... 3
Applications in Research, Gene Therapy
and Biotechnology ................................................................... 5
2. Current Methods for Genotypic and Phenotypic
Knock-Outs in Mammalian Cells ................................................ 9
Antonino Cattaneo and Silvia Biocca
Gene Targeting ............................................................................. 9
Antisense ..................................................................................... 11
Ribozymes ................................................................................... 11
Dominant Negative Mutations ................................................. 13
Ectopic Antibody Expression ................................................... 13
3· Recombinant Antibodies for Ectopic Expression ................... 15
Andrew Bradbury
Antibody Structure .................................................................... 15
Cloning of V Regions for lntracellular Expression ................ 18
Expression of Antihoclies and Antibody Domains ................ 24
Phage Display Antibody Libraries ........................................... 25
Expression of Intracellular Antihoclies ................................... 34
4· Assembly and Folding of Antibodies in Natural
and Artificial Environments ...................................................... 41
Antonino Cattaneo and Silvia Biocca
The Natural Environment:
The Secretory Pathway of B Lymphoid Cells ..................... 42
An Artificial Environment:
Expressing Antibodies in Prokaryotes ............................... 49
Conclusion ................................................................................. 53
5· Protein Sequence Motifs Involved in Intracellular
Trafficking .................................................................................... ;9
Silvia Biocca and Antonino Cattaneo
Sorting in the Secretory Pathway ............................................. 59
Sorting in the Cytosol ................................................................ 67
Signals for Rapid Degradation of Cytosolic Proteins ............. 73
mRNA Targeting ....................................................................... 75
Conclusions ................................................................................ 77
6. Intercellular Immunization ....................................................... 85
Antonino Cattaneo, Patrizia Piccioli and Francesca Ruberti
General Principles ..................................................................... 85
Neuroantibodies: Neurologkai Lesions Created by
Antibody Secreting Neural Cells ......................................... 86
Grafting of Engineered Cells ................................................... 98
Intercellular Immunization for Cell Ablation ...................... 100
7· Intracellular Immunization ..................................................... 105
Silvia Biocca and Antonino Cattaneo
Targeting of Intracellular Antibodies .................................... 105
Assembly, Polding and Stability oflntracellular
Antibodies ............................................................................. 110
Applications of Intracellular Antibodies ................................ 112
Modes of Action of Intracellular Antibodies ......................... 117
8. Gene Therapy and Research Applications oflntrabodies
for Human Infectious Diseases ................................................ 125
Wayne A. Marasco
Inhibition of Postintegration Events
of the HIV -1 Life Cycle ........................................................ 126
Inhibition of Preintegration Events
of the HIV -1 Life Cycle ........................................................ 130
Cellular Targets for Gene Therapy
of HIV -1 Infection and AIDS ............................................... 133
Summary .................................................................................. 134
9· Plantibodies: Immunomodulation and
Immunotherapeutic Potential ................................................. 145
Rosella Franconi, Paraskevi Tavladoraki andEugenio Benvenuto
Plant Transformation Systems ............................................... 145
Antibody Engineering in Plants:
A Chronological Survey ...................................................... 146
Antibody Targeting ................................................................. 147
Antibody Assembly and Secretion ......................................... 157
Immunotherapy Against Plant Diseases ............................... 160
Immunotherapeutic Plantibodies:
Biomedical Applications ..................................................... 163
Perspectives and Conclusions ................................................ 164
10. From Phage Libraries to Intracellular Immunization .......... 173
Nicola Gargano, Luisa Fasulo, Silvia Biocca
and Antonino Cattaneo
Phage Derived Antiborlies for Intracellular
Immunization ....................................................................... 173
Intracellular Antiborlies and the Ye ast
Two Hybrid System ............................................................. 174
Intracellular Libraries in Mammalian Cells .......................... 177
u. Perspectives and Conclusions ................................................. 187
Antonino Cattaneo and Silvia Biocca
Index ....................................................................................................... 191
r.================EDITOR~~====~
Antonino Cattaneo, Ph.D.
Biophysics Sector
International School for Advanced Studies (SISSA)
Trieste, Italy
Chapters 1, 2, 4-7, 10, 11
Silvia Biocca, Ph.D.
Department of Experimental Medicine and Biochemical Seiences
U niversity of Rome "Tor V ergata"
Roma, Italy
s,
Chapters 1, 2, 4, J, 10, 11
r============CONTRIBUTORS============:::::j
Eugenio Benvenuto Nicola Gargano
ENEA, Dipartimento Innovazione MRC Laboratory of Molecular
Divisione Biotecnologie e Biology
Agricoltura Cambridge, U.K.
Roma, Italy Chapter 10
Chapter9
Wayne A. Marasco, M.D., Ph.D.
Andrew Bradbury, Ph.D. Division ofHuman Retrovirology
Biophysics Sector and Infectious Diseases
International School for Advanced Dana-Farber Cancer Institute
Studies (SISSA) Harvard Medical School
Trieste, Italy Boston, Massachusetts, U.S.A.
Chapter 3 ChapterB
Luisa Fasulo Patrizia Piccioli
Biophysics Sector Institute of Neurobiology, CNR
International School for Advanced Roma, Italy
Studies (SISSA) Chapter6
Trieste, Italy
Chapterw Francesca Ruberti
Biophysics Sector
Rosella Franconi International School for Advanced
ENEA, Dipartimento Innovazione Studies (SISSA)
Divisione Biotecnologie e Trieste, Italy
Agricoltura Chapter6
Roma, Italy
Chapter9 Paraskevi Tavladoraki
Universita' Roma Tre,
Dipartimento di Biologia,
Roma, Italy
Chapter9