Human Antibodies to Dendritic Cells Generation: Analysis and Use in Vaccination

156 Pages·1972·1.1 MB·English

by  Annemarie Nicolette Lekkerkerker

H
UMAN ANTIBODIES TO DENDRITIC CELLS
GENERATION, ANALYSIS AND USE IN VACCINATION

H
UMAN ANTIBODIES TO DENDRITIC CELLS
GENERATION, ANALYSIS AND USE IN VACCINATION

HUMANE ANTILICHAMEN TEGEN DENDRITISCHE CELLEN
Ontwikkeling, analyse en gebruik in vaccinatie

(met een samenvatting in het Nederlands)

PROEFSCHRIFT

ter verkrijging van de graad van doctor
aan de Universiteit Utrecht
op gezag van de Rector Magnificus, Prof. dr. W.H. Gispen,
ingevolge het besluit van het College voor Promoties
in het openbaar te verdedigen
op vrijdag 31 mei 2002 des middags te 2.30 uur

door

Annemarie Nicolette Lekkerkerker

geboren op 6 december 1972, te Asse, België
Promotoren Prof. dr. T. Logtenberg
Prof. dr. H. Clevers

Cover: ‘Black Hollyhock with Blue Larkspur’ Georgia O’Keeffe 1929

ISBN: 90-393-3038-7

The studies described in this thesis were performed at the Department of Immunology,
University Medical Center Utrecht, The Netherlands and were financially supported by
Crucell N.V..
When you take a flower in your hand and really look at it, it’s your world for the moment.

Georgia O’Keeffe

CONTENTS

CHAPTER 1
General introduction 9

CHAPTER 2A
Phage antibodies against human dendritic cell subpopulations by 33
flow-cytometry based selection on freshly isolated cells

CHAPTER 2B
Phage antibodies selected on dendritic cells in human tonsil 51

CHAPTER 3
Reactivity profiles of human IgG4 monoclonal antibodies on naturally 69
occurring and cultured dendritic cell subsets

CHAPTER 4
Binding and crosslinking of huMab MatDC 64 prevents the in vitro 97
differentiation of human monocytes into dendritic cells

CHAPTER 5
In vitro antibody-mediated delivery of melanoma tumor antigen MAGE-1 111
to immature dendritic cells results in anti-tumor T and CTL responses
H

CHAPTER 6
Lipid-tagged scFv fragments incorporated in tumor cells mediate 131
receptor-dependent targeting to antigen presenting cells

CHAPTER 7
Summarizing discussion 145

Nederlandse Samenvatting 155

Curriculum Vitae 157

List of Publications 158

Nawoord 159
CHAPTER 1

G
ENERAL INTRODUCTION

GENERAL INTRODUCTION

INTRODUCTION

The life cycle of dendritic cells
Dendritic cells (DCs), first described in 1973 by Steinman and Cohn 1, are now widely
recognized as professional antigen presenting cells (APCs) that play a pivotal role in
directing the immune response. The ability of DCs to process and present a broad variety of
antigens, to which an organism is exposed, is unmatched.
The functioning of the immune system is based on two distinct types of responses, the
innate and the adaptive immune response. The innate immune response, mediated by a
variety of cell types including natural killer (NK) cells and macrophages, forms the first line
of defense towards infections and is regarded as non-specific. The adaptive immune response
depends on the activity of antigen-specific effector T and B cells and is notable for its
exquisite specificity for the inciting antigen and the generation of memory for antigens. DCs
are innate immune cells and acutely activated when a pathogen invades the body, but they
also play a decisive and instructive role in the adaptive immune response that arises 2. Given
that the induction of productive T cell responses depends upon activation of DCs, it follows
that the DC serves as a pivotal interface bridging the innate and adaptive immune system.
Generated in large numbers in the bone marrow every day, precursor DCs circulate in
the blood stream and migrate into tissues, where they reside as sentinels in an immature state,
the prototype being Langerhans cells in the epidermis. Immature DCs have a high phagocytic
and endocytic capacity and constantly sample their environment for antigens at potential sites
of pathogen entry.
Upon encounter of an antigen and the exposure to ‘danger’ signals (pathogens, tissue
damage and local inflammation) immature DCs migrate to the lymphoid organs (Fig. 1).
During this migration, which is regulated by altered expression of chemokine receptors, DCs
undergo the maturation process that is characterized by down-regulation of the capacity to
capture antigen and up-regulation of antigen processing and presentation, and of expression
of co-stimulatory molecules. In the secondary lymphoid organs, mature DCs present antigens
captured in the periphery to resting or naive T cells, inducing an adaptive immune response.
After delivery of their pathogen-related information, the DCs, no longer necessary for the
acute response to antigen, presumably undergo apoptotic cell death and are eliminated.

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