Analysis of C-type lectin receptor induced NF-kappaB signaling:
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| Format: | Thesis Book |
| Language: | English |
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2013
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| Online Access: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20140508-1144877-0-1 Inhaltsverzeichnis |
| Physical Description: | 132 Bl. Ill., graph. Darst. |
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| adam_text | TECHNISCHE UNIVERSITAT MUNCHEN
LEHRSTUHL FUR PROTEOMIK UND BIOANALYTIK
ANALYSIS OF C-TYPE LECTIN RECEPTOR INDUCED NF-KAPPAB SIGNALING
ANDREAS DOMINIKUS STRAFLER
VOLLSTANDIGER ABDRUCK DER VON DER FAKULTAT WISSENSCHAFITSZENTRUM
WEIHENSTEPHAN FUR
ERNAHRUNG, LANDNUTZUNG UND UMWELT DER TECHNISCHEN UNIVERSITAT MUNCHEN
ZUR ERLANGUNG
DES AKADEMISCHEN GRADES EINES
DOKTORS DER NATURWISSENSCHAFTEN (DR. RER. NAT.)
GENEHMIGTEN DISSERTATION.
VORSITZENDER: UNIV.-PROF. DR. DIRK HALLER
PRIIFER DER DISSERTATION: 1. UNIV.-PROF. DR. BERNHARD KIISTER
2. UNIV.-PROF. DR. JIIRGEN RULAND
DIE DISSERTATION WURDE AM 25.06.2013 BEI DER TECHNISCHEN UNIVERSITAT
MUNCHEN EINGEREICHT
UND DURCH DIE FAKULTAT WISSENSCHAFTSZENTRUM WEIHENSTEPHAN FIIR
ERNAHRUNG, LANDNUTZUNG
UND UMWELT AM 12.11.2013 ANGENOMMEN.
HTTP://D-NB.INFO/1053683855
3
TABLE OF CONTENTS
ZUSAMMENFASSUNG 6
SUMMARY 8
1. INTRODUCTION 9
1.1. THE IMMUNE SYSTEM 9
1.1.1. INNATE IMMUNITY 10
1.1.2. ADAPTIVE IMMUNITY 13
1.1.3. EFFECTOR CELLS OF THE INNATE IMMUNE SYSTEM 19
1.1.4. PATTERN RECOGNITION IN INNATE IMMUNITY 24
1.2. THE NUCLEAR FACTOR
K
B PATHWAY 28
1.2.1. NF-
K
B ENGAGEMENT 28
1
.2.2. REL AND I
K
B PROTEIN FAMILIES 29
1.2.3. SIGNAL TRANSDUCTION 32
1.2.4. REGULATION OF TARGET GENE TRANSCRIPTION 35
1
.2.5. RESOLVING NF-
K
B ACTIVITY 37
1.2.6. NF-
K
B SIGNALING IN DISEASE 38
1.3. C-TYPE LECTIN RECEPTORS IN INNATE IMMUNITY 40
1.3.1. DECTIN-1 AND ARCHETYPAL CLR SIGNALING 42
1.3.2. THE CARD9-BCLL0-MALTL SIGNALOSOME 46
1.3.3. CLR-MEDIATED DETECTION OF FUNGAL INVADERS 48
1.4. THE FAMILY OF PROTEIN KINASE C MOLECULES 52
1.4.1. STRUCTURAL CHARACTERISTICS OF THE PKC FAMILY 54
1.4.2. PKC FUNCTION 55
1.4.3. PKCS IN LYMPHOID SIGNALING 57
1.4.3.1. T CELL SPECIFIC ISOFORMS 57
1.4.3.2. B CELL SPECIFIC ISOFORMS 61
1.4.4. PKCS IN INNATE IMMUNITY 64
1.4.5. PROTEIN KINASE C-5 64
1.5. SPECIFIC AIMS OF THIS PROJECT 66
2. MATERIAL AND METHODS 68
2.1. RESEARCH EQUIPMENT 68
TABLE OF CONTENTS 4
2.1.1. LABORATORY APPARATUS 68
2.1.2. MOLECULAR BIOLOGY SUPPLIES 69
2.2. REAGENTS 71
2.2.1. CHEMICALS 71
2.2.2. SOLUTIONS AND BUFFERS 73
2.2.3. MATERIAL AND MEDIA FOR MICROBIOLOGICAL CULTURE 75
2.2.4. MEDIA AND SUPPLEMENTS FOR MAMMALIAN CELL CULTURE 75
2.2.5. ANTIBODIES 75
2.3. METHODS 77
2.3.1. CULTIVATION OF C. ALBICANS 77
2.3.2. MAMMALIAN CELL CULTURE 77
2.3.2.1. BONE MARROW STEM CELL EXTRACTION 77
2.3.2.2. FREEZING BONE MARROW STEM CELLS 78
2.3.2.3. THAWING CELLS 78
2.3.2.4. DENDRITIC CELL CULTURE 78
2.3.3. FUNCTIONAL ASSAYS 79
2.3.3.1. STIMULATION OF DENDRITIC CELLS 79
2.3.3.2. CYTOKINE MEASUREMENT 79
2.3.3.3. LDH-RELEASE ASSAY 80
2.3.3.4. FLOW CYTOMETRY 81
2.3.3.5. INTERNALIZATION OF ZYMOSAN 81
. .2.3.4. PROTEIN ANALYSES 82
2.3.4.1. PRECIPITATION OF PROTEINS FROM CELL CULTURE SUPERNATANTS 82
2.3.4.2. GENERATION OF TOTAL PROTEIN LYSATES 82
2.3.4.3. DETERMINATION OF PROTEIN CONCENTRATION (BRADFORD ASSAY) 83
2.3.4.4. SDS-PAGE AND IMMUNOBLOT ANALYSES 83
2.3.4.5. IMMUNOCHEMICAL DETECTION OF TRANSFERRED PROTEINS 83
2.3.4.6. REMOVAL OF ANTIBODIES (STRIPPING MEMBRANES) 84
3. RESULTS 85
3.1. DECTIN-1 SIGNALING DEPENDS ON PKC ACTIVITY 85
3.1.1. INHIBITOR INFLUENCE ON CELL SURVIVAL 85
3.1.2. INHIBITOR EFFECTS ON PRR SIGNALING 86
3.2. PKC8 IS ESSENTIAL FOR CLR-MEDIATED CYTOKINE PRODUCTION 87
3.2.1. IDENTIFICATION OF THE RELEVANT ISOFORM 87
TABLE OF CONTENTS
5
3.2.2. PHAGOCYTOSIS FUNCTIONS INDEPENDENTLY OF PKC8 IN BMDCS 90
3.2.3. SEVERAL SYK-COUPLED CLRS DEPEND ON PKC8 FOR SIGNALING : 90
3.3. ZYMOSAN STIMULATION TRIGGERS TYROSINE PHOSPHORYLATION OF PK.C8 91
3.4. PKC5 REGULATES DECTIN-1 -MEDIATED NF-KB SIGNALING 92
3.4
.1
. NF-
K
B SIGNALING IS COMPROMISED IN PKC8-DEFICIENT BMDCS 92
3.4.2. CARD9 IS ACTIVATED INDEPENDENTLY OF PKC8 94
3.5. PKC8 ACTIVATES TAK1 VIA CARD9-BCLL0 COMPLEX FORMATION 94
3.6. PKC5 IS ESSENTIAL FOR INNATE ANTI-FUNGAL IMMUNE DEFENSE 97
4. DISCUSSION 100
4.1. DECTIN-L-SYK SIGNALING SPECIFICALLY REQUIRES THE PKC8 ISOFORM 100
4.2. SIGNALING THROUGH PKC8 IS CRITICAL FOR CARD9 AND TAK1 ENGAGEMENT
101
4.3. PKC8 SELECTIVELY REGULATES DECTIN-1-SIGNALING OUTCOMES 102
4.4. PKC8 FUNCTIONS AS A GENERAL MEDIATOR OF CLR SIGNALING 104
4.5. PKC8 IN HOST DEFENSE AGAINST NON-FUNGAL PATHOGENS 105
4.6. LINKING INNATE TO ADAPTIVE IMMUNITY VIA PKC8 105
BIBLIOGRAPHY 107
LIST OF ABBREVIATIONS 122
LIST OF FIGURES AND TABLES 129
PUBLICATIONS 130
ACKNOWLEDGEMENTS 131
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| spelling | Straßer, Andreas Dominikus Verfasser aut Analysis of C-type lectin receptor induced NF-kappaB signaling Andreas Dominikus Straßer 2013 132 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2014 Zsfassung in dt. Sprache (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20140508-1144877-0-1 http://mediatum.ub.tum.de/node?id=1144877 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20140508-1144877-0-1 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027341733&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Straßer, Andreas Dominikus Analysis of C-type lectin receptor induced NF-kappaB signaling |
| subject_GND | (DE-588)4113937-9 |
| title | Analysis of C-type lectin receptor induced NF-kappaB signaling |
| title_auth | Analysis of C-type lectin receptor induced NF-kappaB signaling |
| title_exact_search | Analysis of C-type lectin receptor induced NF-kappaB signaling |
| title_full | Analysis of C-type lectin receptor induced NF-kappaB signaling Andreas Dominikus Straßer |
| title_fullStr | Analysis of C-type lectin receptor induced NF-kappaB signaling Andreas Dominikus Straßer |
| title_full_unstemmed | Analysis of C-type lectin receptor induced NF-kappaB signaling Andreas Dominikus Straßer |
| title_short | Analysis of C-type lectin receptor induced NF-kappaB signaling |
| title_sort | analysis of c type lectin receptor induced nf kappab signaling |
| topic_facet | Hochschulschrift |
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