Crosstalk between autoreactive T cells and alveolar type II epithelial cells in inflammation and tolerance [Elektronische Ressource] / von Marcus Gereke

Andrea Rea

Sujets

• Gesundheit

Informations

• Publié par: technische_universitat_carolo-wilhelmina_zu_braunschweig
• Publié le: 01 janvier 2007
• Nombre de lectures: 36
• Langue: Deutsch
• Poids de l'ouvrage: 3 Mo

Extrait

Crosstalk between autoreactive T cells and
alveolar type II epithelial cells in
inflammation and tolerance




Von dem Fachbereich für Biowissenschaften und Psychologie
der Technischen Universität Carolo-Wilhelmina
zu Braunschweig
zur Erlangung des Grades eines
Doktors der Naturwissenschaften
(Dr.rer.nat.)
genehmigte
D i s s e r t a t i o n


von

Marcus Gereke
aus
Bad Harzburg




























von: Marcus Gereke
aus: Bad Harzburg
1. Referent: Prof. Dr. J. Wehland
2. Referent: Prof. Dr. S. Dübel
eingereicht am: 10.10.2005
mündliche Prüfung
(Disputation) am: 23.02.2006
Druckversion: 2007 Vorveröffentlichungen der Dissertation


Teilergebnisse aus dieser Arbeit wurden mit Genehmigung des Fachbereichs der
Biowissenschaften und Psychologie, vertreten durch den Mentor der Arbeit, in
folgenden Beiträgen vorab veröffentlicht:



Publikationen:

Bruder D, Westendorf AM, Geffers R, Gruber AD, Gereke M, Enelow RI, Buer J.
CD4 T Lymphocyte-mediated lung disease: steady state between pathological and
tolerogenic immune reactions. Am J Respir Crit Care Med. 2004 Dec 1; 170
(11):1145-52.


Tagungsbeiträge:

Gereke M, Westendorf AM, Buer J, and Bruder D. Induction of regulatory T cells
by alveolar self antigen expression. (Poster). Keystone Symposia - Translational
Medicine in Autoimmunity, Big Sky, Montana, USA, 2005.

Gereke M, Prettin S, Gröbe L, Kasper M, Buer J, and Bruder D. The role of type II
alveolar epithelial cells for the induction of T cell tolerance and immune regulation.
(Vortrag). Joint annual meeting of the German and Scandinavian Societies for
Immunology, Kiel, Germany, 2005.







IZusätzliche Tagungsbeiträge:

Bruder D, Westendorf AM, Geffers R, Gereke M, Gruber A, and Buer J. Toleranz
+versus Autoimmunität: Autoreaktive CD4 T-Zellen in der Pathogenese chronisch
entzündlicher Erkrankungen der Lunge. (Vortrag). Tagung der Sektion Zellbiologie
der Deutschen Gesellschaft für Pneumologie, Magdeburg, Germany, 2003.

Gereke M, Westendorf AM, Geffers R, Buer J, Bruder D. Alveolar self antigen
expression leads to the induction of regulatory T cells. (Vortrag). Joint meeting of the
Dutch and German Societies for Immunology, Maastricht, Netherlands, 2004.

Bruder D, Gereke M, Westendorf AM, Geffers R, Enelow RI, Buer J. Induction of
thregulatory T cells due to chronic self antigen stimulation in the lung. (Poster). 12
International Congress of Mucosal Immunology, Boston, USA, 2005.













IITABLE OF CONTENT
Contents


Vorveröffentlichungen der Dissertation I
Contents III
List of Figures VI Tables VIII

CHAPTER I
Introduction 1
1 The mucosal immune system 2
2 The respiratory tract in context of immunity 5
2.1 Immune cells and structures of the lung 5
2.1.1 Macrophages 6
2.1.2 Dendritic cells 7
2.1.3 Lymphocytes 8
2.1.4 Airway epithelium 9
2.1.5 Alveolar epithelium – Type II and Type I
alveolar epithelial cells 11
2.2 The alveolar type II epithelial cell as an integrative unit of the alveolus 13
2.3 rs the source of alveolar surfactant 16
2.3.1 Immunoregulatory functions of surfactant proteins 18
2.3.2 Collectin structure 18
2.3.3 Collectin regulation of immune cells 20
3 Autoimmunity 24
4 Regulatory T lymphocytes 27
+ +4.1 Naturally occurring CD4 CD25 regulatory T cells 28
4.2 Adaptive regulatory T cells 30
4.3 Mechanism of suppression 31
5 SPC-HA transgenic mouse model 33

CHAPTER II
+Results Part I - Characterization of autoreactive CD4 T cells
from SPC-HA x TCR-HA mice 38
1 Aims of the study 39
2 Results 40
+2.1 HA-specific CD4 T cells are present in the periphery
of SPC-HA x TCR-HA mice 40

IIITABLE OF CONTENT
+2.2 HA-specific CD4 T lymphocytes from SPC-HA x TCR-HA mice
have an activated phenotype 42
+ +2.3 Marker genes for regulatory T cells are expressed in 6.5 CD4
lung lymphocytes isolated from SPC-HA x TCR-HA double
transgenic mice 47
+2.4 Progressive infiltration of HA-specific CD4 T cells in the lung
of SPC-HA mice after adoptive transfer 48
+ + + +2.5 Adoptive transfer of 6.5 CD4 and 6.5 CD4 depleted of
+CD25 regulatory T cells into SPC-HA transgenic mice 50
+ +2.6 Adoptive transfer of 6.5 CD4 T cells in SPC-HA x TCR-HA
double transgenic mice 52

CHAPTER II
Results Part II - Characterization of self-antigen expressing alveolar type II
epithelial cell from SPC-HA x TCR-HA mice 54
3 Aims of the study 55
4 Results 57
4.1 Isolation of murine alveolar type II epithelial cells 57
4.2 Global gene expression profiling of murine alveolar type II
epithelia cels 61
4.3 Analysis of genes differentially expressed in AECII from
SPC-HA x TCR-HA mice double transgenic mice and
SPC-HA transgenic mice 65
4.4 Antigen presentation in the lung 69
4.5 Presentation of AECII expressed self-antigen mediated
by Dendritc ells 70
4.6 Peritoneal exudate cells (PEC) as antigen presenting cells
can not mediate self-antigen presentation to autoreactive T cells 72
4.7 The antigen presenting capacity of alveolar type II epithelial cells 73
4.8 Reduced stimulatory capacity of AECII from SPC-HA x TCR-HA
double transgenic mice 76
4.9 Analysis of hemagglutinin expression in AECII SPC-HA x TCR-HA
77 double transgenic mice and SPC-HA transgenic mice
+4.10 The impact of AECII conditioned media on CD4 T cell proliferation 78
4.11 Differences in the surfactant protein expression in AECII from
SPC-HA x TCR-HA double transgenic mice and SPC-HA
79 transgenic mice
+4.12 The effect of AECII/CD4 T cell coculture on the T cell
phenotype and function 81


IVTABLE OF CONTENT
CHAPTER III
Discussion and Summary 85
1 Discussion 86
2 Summary 100

CHAPTER IV
Materials and Methods 102
1 Materials and Methods 103
1. Mice 103
1.2Preparation of lymphocyte populations 103
1.3 Primary Type II Pneumocytes Preparation 104
1.4 Antibodies and flow cytometry 105
1.5 Carboxyfluorescein diacetate, succinimidyl ester (CFSE)
labeling of lymphocytes 105
1.6 Adoptive transfer 106
1.7 Histology 106
1.8 Proliferation assay and cocultures experiments 107
1.9 Inhibition assay 107
1.10 Expression analysis by RT-PCR and real time RT-PCR 108
1.11 DNA microarray hybridization and analysis 110
1.12 Immunofluorescence 111
1.13Isolation of Dendritic cells (DC) 111

CHAPTER V
Abbreviations and References 112
1 Abbreviations 113
2 References 115

VList of Figures

Figure 1: The basic structure of BALT and their distribution of lymphocyte
subsets and macrophages. 4
Figure 2: Schematic structure of airway and alveolus and their host-defence
mechanisms. 13
Figure 3: Collectin and C1q structure. 19
Figure 4: Collectin receptors. 23
Figure 5: Schematic representation of T cell development. 26
Figure 6: Two classes of regulatory T cells can be envisioned. 32
Figure 7: Targeted expression of hemagglutinin in the lung. 33
Figure 8: Immunohistology analysis of lung from SPC-HA x TCR-HA double
transgenic mouse compared to SPC-HA transgenic mouse. 34
+Figure 9: Increased number of HA-specific CD4 T cells in the lung of
SPC-HA x TCR-HA mice. 35
+ +Figure 10: Cluster analysis of genes differentially expressed in 6.5 CD4
T cells isolated from lungs and spleens of diseased
SPC-HA x TCR-HA as well as healthy TCR-HA mice. 36
+Figure 11: HA-specific CD4 T cells are present in periphery of
SPC-HA x TCR-HA mice. 40
+Figure 11 (continued): HA-specific CD4 T cells are present in periphery of
SPC-HA x TCR-HA mice. 41
+Figure 11 (continued): HA-specific CD4 T cells are present in periphery of
SPC-HA x TCR-HA mice. 42
+Figure 12: Activation/memory