Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP): modulators of the immune system
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| Format: | Thesis Book |
| Language: | English German |
| Published: |
2014
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in engl. und dt. Sprache |
| Physical Description: | 178 S. Ill., graph. Darst. |
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| adam_text | SOLUBLE CD83 (SCD83) AND
THYMIC STROMAL LYMPHOPOIETIN (TSLP):
MODULATORS OF THE IMMUNE SYSTEM
DER NATURWISSENSCHAFTLICHEN FAKULTAT
DER FRIEDRICH-ALEXANDER-UNIVERSITAT ERLANGEN-NURNBERG
ZUR ERLANGUNG DES DOKTORGRADES DR. RER. NAT.
VORGELEGT VON
JENNY ECKHARDT
AUS MARBURG AN DER LAHN
HTTP://D-NB.INFO/1053634986
TABLE OF CONTENTS
1 SUMMARY 1
1.1 SUMMARY: SOLUBLE CD83 AMELIORATES EXPERIMENTAL COLITIS 1
1.2 ZUSAMMENFASSUNG: LOSLICHES CD83 SCHWACHT DNBS INDUZIERTE COLITIS AB
2
1.3 SUMMARY: THYMIC STROMAL LYMPHOPOIETIN (TSLP) DEFICIENCY AMELIORATES
EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) 4
1.4 ZUSAMMENFASSUNG: ABGESCHWACHTE EXPERIMENTELLE AUTOIMMUNE
ENZEPHALOMYELITIS (EAE) BEI THYMIC STROMAL LYMPHOPOIETIN
(TSLP)-DEFIZIENZ 6
2 INTRODUCTION 9
2.1 THE IMMUNE SYSTEM 9
2.1.1 THE INNATE IMMUNE SYSTEM 10
2.1.2 THE ADAPTIVE IMMUNE SYSTEM 11
2.2 CELLS OF THE IMMUNE SYSTEM 12
2.2.1 DENDRITIC CELLS 12
2.2.2 CYTOKINES INFLUENCE THE DIFFERENTIATION OF CD4* T CELLS 14
2.2.3 DIFFERENT FUNCTIONS OF CD4* T CELLS 16
2.3 CYTOKINES AND SOLUBLE FACTORS ARE MODULATORS OF THE IMMUNE SYSTEM 18
2.3.1 IMMUNE - MODULATING MOLECULES: SCD83 AND TSLP 18
2.3.2 CD83 19
2.3.3 DIFFERENT FUNCTIONS OF SOLUBLE AND MEMBRANE-BOUND CD83 20
2.3.4 THYMIC STROMAL LYMPHOPOIETIN (TSLP) 27
2.4 AUTOIMMUNE DISEASES 34
2.4.1 INFLAMMATORY BOWEL DISEASE (IBD) 34
2.4.2 CYTOKINES IN IBD 35
2.4.3 DNBS-INDUCED COLITIS: A MOUSE MODEL OF CROHN S DISEASE (CD) 37
2.4.4 MULTIPLE SCLEROSIS (MS) 37
2.4.5 EXPERIMENTAL AUTOIMMUNE ENCEPHALITIS (EAE) 38
2.4.6 CYTOKINE KNOCKOUT MICE IN EAE 40
2.4.7 THE ROLE OF CHEMOKINES AND CHEMOKINE RECEPTORS IN EAE 41
2.4.8 MURINE WHOLE-ORGAN IMMUNE CELL POPULATIONS REVEALED BY
MULTI-EPITOPE-
LIGAND CARTOGRAPHY (MELC) 42
2.5 FORMULATION OF THE THESIS 46
2.5.1 THESIS
1: SOLUBLE CD83 AMELIORATES EXPERIMENTAL COLITS 46
2.5.2 THESIS 2: TSLP DEFICIENCY AMELIORATES EAE 47
3 MATERIALS AND METHODS 49
3.1 MATERIAL 49
3.1.1 CONSUMABLES 49
3.1.2 LABORATORY EQUIPMENT 49
3.1.3 CHEMICALS 50
3.1.4 BUFFERS 51
3.1.5 DNA OLIGONUCLEOTIDES FOR POLYMERASE CHAIN REACTION (PCR) 52
3.1.6 ANTIBODIES AND REAGENTS FOR FACS 55
3.1.7 ANTIBODIES FOR HISTOLOGY 56
3.1.8 CELL MEDIA AND ADDITIVES 57
3.1.9 CYTOKINES, CHEMOKINES, PEPTIDES, PROTEINS AND CELL ACTIVATING
AGENTS 57
3.1.10 ENZYMES 58
3.1.11 KITS 58
3.1.12 RADIOACTIVE CHEMICALS 59
3.1.13 MICE 59
3.1.14 SOFTWARE 59
3.1.15 STATISTICAL ANALYSES 59
3.2 METHODS 60
3.2.1 PREPARATION OF TOTAL RNA AND CDNA SYNTHESIS 60
3.2.2 REALTIME RT-PCR 60
3.2.3 PREPARATION OF MONONUCLEAR CELL (MNC) SUSPENSIONS 61
3.2.4 GENERATION OF BONE MARROW-DERIVED DENDRITIC CELLS (BM-DCS) 62
3.2.5 STIMULATION OF BONE MARROW-DERIVED DENDRITIC CELLS (BM-DCS) 63
3.3 SPECIFIC METHODS USED IN CHAPTER 1: MURINE WHOLE ORGAN IMMUNE CELL
POPULATIONS REVEALED BY MELC 64
3.3.1 SAMPLE PREPARATION FOR MELC 64
3.3.2 GENERATION OF PHASE CONTRAST AND FLUORESCENCE IMAGES 64
3.3.3 MELC DATA ANALYSIS 65
3.4 SPECIFIC METHODS USED IN CHAPTER 2: SOLUBLE CD83 AMELIORATES
EXPERIMENTAL
COLITIS 65
3.4.1 ANIMALS 65
3.4.2 INDUCTION OF EXPERIMENTAL COLITIS 65
3.4.3 IMMUNOSUPPRESSION BY SCD83 65
3.4.4 INDOLEAMINE 2, 3-DIOXYGENASE (IDO) INHIBITION BY
1-METHYL-TRYPTOPHAN (1-
MT) PELLET IMPLANTATION 66
3.4.5 MACROSCOPIC ASSESSMENT 66
3.4.6 HISTOLOGICAL ASSESSMENT 66
3.4.7 SCD83 DETECTION ELISA 68
3.4.8 FLOW CYTOMETRY 69
3.4.9 DC MIGRATION ASSAY 70
3.4.10 B-GLUCORONIDASE ASSAY 71
3.5 SPECIFIC METHODS USED IN CHAPTER 3: TSLP DEFICIENCY AMELIORATES
EAE 71
3.5.1 ANIMALS 71
3.5.2 GENOMIC DNA (GDNA) PURIFICATION 72
3.5.3 MOUSE TAIL POLYMERASE CHAIN REACTION (PCR) 72
3.5.4 INDUCTION OF EXPERMINTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) 73
3.5.5 RATING OF CLINICAL EAE SYMPTOMS 73
3.5.6 REMOVAL OF BLOOD CONTAMINATIONS FROM CNS 74
3.5.7 NEUROPATHOLOGY 74
3.5.8 TREATMENT OF DEREG MICE WITH DIPHTERIA TOXIN 75
3.5.9 FLOW CYTOMETRY ANALYSES...; 75
3.5.10 ANALYSES OF BM-DCS 77
3.5.11 CYTOMETRIC BEAD ARRAY (CBA) OF SERUM 78
3.5.12 T CELL PROLIFERATION AND RESTIMULATION ANALYSES 78
4 RESULTS 80
4.1 CHAPTER 1: MURINE WHOLE ORGAN IMMUNE CELL POPULATIONS REVEALED BY
MELC . 80
4.1.1 MELC TECHNOLOGY FOR THE DETECTION OF MURINE LEUKOCYTES 80
4.1.2 MELC FOR THE
CHARACTERIZATION OF LYMPHOID ORGANS 80
4.1.3 MELC FOR THE
DETERMINATION OF THE ACTIVATION STATE 83
4.1.4 CHARACTERIZATION OF INFLAMMATORY INFILTRATES IN EAE USING MELC 85
4.2 CHAPTER 2: SOLUBLE CD83 AMELIORATES EXPERIMENTAL COLITIS 88
4.2.1 CD83 IS EXPRESSED IN THE GASTROINTESTINAL (GL) TRACT AND IN
MESENTERIAL LYMPH
NODES (MLN) 88
4.2.2 SCD83 TREATMENT AMELIORATES SYMPTOMS OF DNBS-INDUCED COLITIS 94
4.2.3 SCD83 TREATMENT INHIBITS INFLAMMATION OF THE COLON 97
4.2.4 SCD83 ENHANCES IDO EXPRESSION IN MLN 99
4.2.5 INHIBITION OF IDO INTERFERES WITH SCD83-INDUCED PROTECTIVE EFFECT
105
4.3 CHAPTER 3: TSLP DEFICIENCY AMELIORATES EAE 108
4.3.1 DELAYED AND AMELIORATED EAE DEVELOPMENT IN TSLP KO MICE 108
4.3.2 TSLP DEFICIENCY DOES NOT ALTER IMMUNE CELL HOMEOSTASIS IN HEALTHY
MICE. 110
4.3.3 BM-DCS FROM TSLP KO AND WT MICE SHOW NO PHENOTYPIC DIFFERENCES 113
4.3.4 DCS OF TSLP KO MICE DO NOT SHOW
A REDUCED MATURATION DURING EAE 114
4.3.5 IN TSLP KO MICE TH2 RESPONSES ARE NOT REDUCED DURING EAE 115
4.3.6 REDUCED CNS INFLAMMATION IN TSLP KO MICE AT DAY 11 OF EAE 117
4.3.7 REDUCED NUMBERS OF THL CELLS IN CNS OF TSLP KO MICE AT DAY 11 OF
EAE... 119
4.3.8 ENCEPHALITOGENIC T CELLS FROM TSLP KO MICE SHOW
A REDUCED ACTIVATION AT
DAY 11 OF EAE 120
4.3.9 AT THE LATE PHASE OF DISEASE T CELLS
FROM TSLP KO AN WT MICE SHOW NO
DIFFERENCES IN ACTIVATION 121
4.3.10 LYMPH NODE CELLS FROM TSLP KO MICE SHOW A REDUCED ALLOGENEIC
RESPONSE IN
THE MIXED LYMPHOCYTE REACTION (MLR) 123
4.3.11 TSLP CAN DIRECTLY ACTIVATE T CELLS 124
DISCUSSION 125
5.1 CHAPTER 1: MURINE WHOLE ORGAN IMMUNE CELL POPULATIONS REVEALED BY
MELC125
5.2 CHAPTER 2: SOLUBLE CD83 AMELIORATES EXPERIMENTAL COLITIS 128
5.2.1 SOLUBLE CD83 HAS IMMUNOSUPPRESSIVE FUNCTIONS 128
5.2.2 CD83 IS EXPRESSED IN THE GL TRACT UNDER PHYSIOLOGIC CONDITIONS 128
5.2.3 COLON CELLS FROM DNBS-TREATED MICE RELEASE SCD83 129
5.2.4 SCD83 AMELIORATES DNBS-INDUCED COLITIS 130
5.2.5 SCD83 DECREASES INFLAMMATORY REACTIONS IN THE COLON 130
5.2.6 SCD83 NORMALIZES CELL COMPOSITION AND CYTOKINE SECRETION OF MLN
131
5.2.7 SCD83 INDUCES IDO EXPRESSION IN MLN 132
5.2.8 THE IDO INHIBITOR 1-MT ABROGATES THE PROTECTIVE EFFECTS OF SCD83
132
5.2.9 THE INFLUENCE OF SCD83 ON APOPTOSIS 133
5.2.10 THE INFLUENCE OF SCD83 ON CELL MIGRATION 133
5.2.11 THE INFLUENCE OF SCD83 ON CELL PROLIFERATION 134
5.2.12 CONCLUSION 134
5.3 CHAPTER 3: TSLP DEFICIENCY AMELIORATES EAE 135
5.3.1 LEUKOCYTE NUMBERS IN NAIVE TSLP KO MICE ARE NORMAL 135
5.3.2 TSLP IS NOT ESSENTIAL FOR TREG DEVELOPMENT IN MICE 135
5.3.3 TSLP DEFICIENCY HAS NO EFFECT ON DC MATURATION OR FUNCTION 136
5.3.4 TSLP EXPRESSION IS INDUCED BY LPS AND TNFA 137
5.3.5 TSLP DEFICIENCY DOES NOT LED TO REDUCED TH2 RESPONSES DURING EAE
137
5.3.6 TSLP KO MICE SHOW AN AMELIORATED FORM OF EAE 137
5.3.7 REDUCED INFLAMMATORY INFILTRATES IN THE CNS OF TSLP KO MICE 138
5.3.8 ENCEPHALITOGENIC T CELLS FROM KO MICE SHOW A REDUCED ACTIVATION
139
5.3.9 TSLP ACTIVATES
T CELLS
EITHER DIRECTLY OR INDIRECTLY VIA MYDCS 139
5.3.10 LN CELLS FROM TSLP KO MICE SHOW
A REDUCED ALLOGENEIC RESPONSE 140
5.3.11 T CELLS
FROM TSLP KO MICE SHOW A REDUCED MOG
3
S-5S
-SPECIFIC RESPONSE.... 142
5.3.12 CONCLUSION 143
6 REFERENCES 144
7 ABBREVIATIONS .-. 173
8 DANKSAGUNG 175
9 LEBENSLAUF. 176
10 PUBLIKATIONEN 178
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| spelling | Eckhardt, Jenny 1979- Verfasser (DE-588)1053060807 aut Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system vorgelegt von Jenny Eckhardt 2014 178 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in engl. und dt. Sprache Erlangen-Nürnberg, Univ., Diss., 2014 Immunmodulation (DE-588)4315157-7 gnd rswk-swf Cytokine (DE-588)4205701-2 gnd rswk-swf Antigen CD83 (DE-588)4767577-9 gnd rswk-swf Immunsystem (DE-588)4026643-6 gnd rswk-swf Chronische Darmentzündung (DE-588)4273697-3 gnd rswk-swf Encephalomyelitis (DE-588)4152136-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Chronische Darmentzündung (DE-588)4273697-3 s Encephalomyelitis (DE-588)4152136-5 s Immunsystem (DE-588)4026643-6 s Immunmodulation (DE-588)4315157-7 s Cytokine (DE-588)4205701-2 s Antigen CD83 (DE-588)4767577-9 s DE-188 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027395880&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Eckhardt, Jenny 1979- Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system Immunmodulation (DE-588)4315157-7 gnd Cytokine (DE-588)4205701-2 gnd Antigen CD83 (DE-588)4767577-9 gnd Immunsystem (DE-588)4026643-6 gnd Chronische Darmentzündung (DE-588)4273697-3 gnd Encephalomyelitis (DE-588)4152136-5 gnd |
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| title | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system |
| title_auth | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system |
| title_exact_search | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system |
| title_full | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system vorgelegt von Jenny Eckhardt |
| title_fullStr | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system vorgelegt von Jenny Eckhardt |
| title_full_unstemmed | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) modulators of the immune system vorgelegt von Jenny Eckhardt |
| title_short | Soluble CD83 (sCD83) and thymic stromal lymphopoietin (TSLP) |
| title_sort | soluble cd83 scd83 and thymic stromal lymphopoietin tslp modulators of the immune system |
| title_sub | modulators of the immune system |
| topic | Immunmodulation (DE-588)4315157-7 gnd Cytokine (DE-588)4205701-2 gnd Antigen CD83 (DE-588)4767577-9 gnd Immunsystem (DE-588)4026643-6 gnd Chronische Darmentzündung (DE-588)4273697-3 gnd Encephalomyelitis (DE-588)4152136-5 gnd |
| topic_facet | Immunmodulation Cytokine Antigen CD83 Immunsystem Chronische Darmentzündung Encephalomyelitis Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027395880&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT eckhardtjenny solublecd83scd83andthymicstromallymphopoietintslpmodulatorsoftheimmunesystem |