B-cell Development and Primary Antibody Deficiencies - RePub

196 Pages·2007·7.8 MB·English

B-cell Development and Primary Antibody Deficiencies
B-celontwikkeling en primaire antistofdeficiënties
ISBN-13: 978-90-73436-76-3
“The quest of the B cell”
Developing B cells have a long way to go before they are ready to encounter disease-
causing pathogens. In order to make it a successful journey, multiple factors are re-
quired, without which the guest will fail. The ultimate dream of the B cell is to fight a
pathogen and become an antigen-experienced B cell that can generate a fast and strong
immune response, which contributes to antigen-specific immunity.
No part of this thesis may be reproduced or transmitted in any form by any means, electronic or
mechanical, including photocopying, recording or any information storage and retrieval system,
without permission in writing from the publisher (M.C. van Zelm, Department of Immunology,
Erasmus MC, P.O. Box 2040, 3000 CA Rotterdam, the Netherlands).
B-cell Development and Primary Antibody Deficiencies
B-celontwikkeling en primaire antistofdeficiënties
PROEFSCHRIFT
ter verkrijging van de graad van doctor
aan de Erasmus Universiteit Rotterdam
op gezag van de rector magnificus
Prof.dr. S.W.J. Lamberts
en volgens besluit van het College voor Promoties.
De openbare verdediging zal plaatsvinden op
woensdag 20 juni 2007 om 13.45 uur
door
Menno Cornelis van Zelm
geboren te Utrecht
PROMOTIECOMMISSIE
Promotoren: Prof.dr. J.J.M. van Dongen
Prof.dr. R. de Groot
Overige leden: Prof.dr. H. Hooijkaas
Prof.dr. C. Murre
Prof.dr. C.J.M. van Noesel
Copromotor: Dr. M. van der Burg
The studies described in this thesis were
performed at the Department of Immunology,
Erasmus MC, Rotterdam, the Netherlands and
partly at the division of Biological Sciences,
University of California San Diego, La Jolla,
CA, USA.

The studies were financially supported by ‘Sophia Kinderziekenhuis Fonds’
(SKF; grant 349), and partly by Ter Meulen Fund - Royal Netherlands Academy of Arts
and Sciences (KNAW), Trustfonds - Erasmus University Rotterdam and the Dutch Society
for Immunology (NVvI).
The printing of the thesis was supported by Erasmus University Rotterdam, J.E. Jurriaanse
Stichting, Stichting Kind & Afweer, Baxter B.V., Dr. Ir. Van de Laar Stichting,
Beckman Coulter Nederland B.V. and BD Biosciences.
Illustrations : W. Marieke Comans-Bitter
Printing : Ridderprint Offsetdrukkerij B.V., Ridderkerk
Cover : Frans Tutert
Lay-out : Wendy Netten
Voor mijn ouders
B-cell Development and Primary Antibody Deficiencies
B-celontwikkeling en primaire antistofdeficiënties
CONTENTS
Chapter I General Introduction 9

Chapter II Ig Gene Rearrangement Steps are Initiated in Early 41
Human Precursor-B-cell Subsets and Correlate with
Specific Transcription Factor Expression
J Immunol 2005;175(9):5912-22

Chapter III 3D-Architecture of the Immunoglobulin Heavy Chain Locus 67
Submitted
Chapter IV Gross Deletions involving IGHM, BTK or Artemis: 93
A model for Genomic Lesions Mediated by Transposable
Elements
Submitted
Chapter V An Antibody-Deficiency Syndrome Due to Mutations in 115
the CD19 Gene
N Engl J Med 2006;354(18):1901-12
Chapter VI Replication History of B lymphocytes Reveals 133
Homeostatic Proliferation and Extensive Antigen-
induced B-cell Expansion
J Exp Med 2007;204(3):645-55
Chapter VII General Discussion 153
Chapter VIII Appendices 167
1 PCR primers and TaqMan probes for quantification of Ig gene 168
rearrangements
2 PCR primers for GeneScan analysis of complete IGH, IGK and 170
IGL rearrangements
3 PCR primers and TaqMan probes for quantification of expression levels of 171
16 candidate genes for induction of Ig gene rearrangements
4 Full width at half the maximum (FWHM) values of the point spread function 172
for the indicated colors
5. Chromatic shifts 172
6 Resolution equivalents before and after application of chromatic corrections 172
7 IGHM gross deletion primers for deletion-scanning and breakpoint PCR 173
8 BTK gross deletion primers for deletion-scanning and breakpoint PCR 177
9 Artemis gross deletion primers for deletion-scanning and breakpoint PCR 178
10 PCR primers for amplification and sequencing of CD19 coding regions from 179
genomic DNA
11 PCR primers and TaqMan probe for quantification of CD19 transcripts 179
12 PCR primers and TaqMan probes for KREC and IgκREHMA assays 180
Abbreviations 181

Summary 183

Samenvatting 186

Dankwoord 190

Curriculum Vitae 192
Publications 194

I
General Introduction
B-cell Development and Primary Antibody Deficiencies
The immune system of higher vertebrates, such as humans, is a complex interplay of cells
and protein products that protects the body from invading micro-organisms. The first line of
defense is formed by the skin and mucous membranes. The second line consists of proteins
(e.g. complement) and immune cells (e.g. macrophages, granulocytes, natural killer (NK)
cells) that are part of the innate immune system. T and B lymphocytes are part of the specific
adaptive immune system: the third line of defense. These lymphocytes adapt their response
during an infection to improve recognition of the pathogen and this improved response is then
retained after the pathogen has been eliminated in the form of an immunological memory.
Classically, the cells of the innate immune system were thought to react a-specifically, whereas
the adaptive immune cells carried out a specific immune response. However, more recently
innate immune cells were found to carry evolutionarily conserved receptors, named Toll-like
receptors (TLRs). These receptors play a crucial role in early host defense against invading
pathogens and are specific enough to discriminate self and pathogens.
B lymphocytes contribute to the immune system by production of antigen-specific
antibodies (immunoglobulins; Ig). The antibody response to a pathogen is the classical
example to demonstrate the enormous capacity and specificity of the adaptive immune response
(Figure 1). Similar to most immune cells, B lymphocytes are generated throughout life from
hematopoietic stem cells in bone marrow (Figure 2). Every newly generated B-lymphocyte
carries a specific Ig receptor, which has been generated during differentiation in bone marrow
Primary response Secondary response
104
103 lag
phase
m)
u102
er
s
ml 101
g/

y (100
d response to
o
b antigen A
nti10-1
A
10-2
response
to antigen B
10-3
4 8 12 16 20 64 68 72
antigen A Days antigens
A + B
Figure 1. The course of a typical immune response.
Upon first encounter with antigen A, an immune response is initiated and antibodies against antigen A are produced
after a lag phase of several days. The antibody levels reach a plateau and then decrease. When the animal is later
challenged with antigen A, a very rapid and intense response occurs. The response to antigen B at the same time of the
booster with antigen A, resembles the initial and primary response to antigen A. This illustrates the antigen-specific
build up of memory. Figure adapted from Janeway et al. Immunobiology, 2001, Garland Science.
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