Mycobacterial cord factor mediated activation of murine macrophages: analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | German English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIX, 118, III S. Ill., graph. Darst. |
Internformat
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| 100 | 1 | |a Schoenen, Hanne Gunda |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Mycobacterial cord factor mediated activation of murine macrophages |b analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |c vorgelegt von Hanne Gunda Schoenen |
| 246 | 1 | 1 | |a Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor |
| 264 | 1 | |c 2013 | |
| 300 | |a XIX, 118, III S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |a Erlangen-Nürnberg, Univ., Diss., 2013 | ||
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Datensatz im Suchindex
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| adam_text | IMAGE 1
TABLE OF CONTENT
TABLE OF CONTENT
TABLE OF CONTENT I
SUMMARY VI
ZUSAMMENFASSUNG VIII
ABBREVIATIONS X
INDEX OF FIGURES XVI
INDEX OF TABLES XIX
1. INTRODUCTION 1
1.1 THE IMMUNE SYSTEM 1
1.2 THE MACROPHAGE 1
1.2.1 MACROPHAGE DIFFERENTIATION 2
1.2.2 IMMUNOLOGICAL EFFECTOR FUNCTIONS OF MACROPHAGES 2
1.2.3 TRANSCRIPTION FACTORS REGULATING MACROPHAGE EFFECTOR FUNCTIONS 4
1.2.3.1 CONSTITUTIVELY EXPRESSED TRANSCRIPTION FACTORS 4
1.2.3.2 INDUCIBLE TRANSCRIPTION FACTORS 6
1.3 RECOGNITION OF MICROBIAL UGANDS 9
1.3.1 TOLL-UKE RECEPTORS 9
1.3.1.1 TLRS AND THEIR LIGANDS 10
1.3.1.2 INTRACELLULAR SIGNALING INDUCED BY TLRS 10
1.3.2 C-TYPE LECTIN RECEPTORS 12
1.3.2.1 CLRS AND THEIR LIGANDS 12
1.3.2.2 INTRACELLULAR SIGNALING INDUCED BY CLRS 13
1.3.3 THE MYCOBACTERIAL CELL WALL AND RECOGNITION OF SELECTED COMPONENTS
15
1.3.3.1 TLR LIGANDS DERIVED FROM THE MYCOBACTERIAL CELL WALL 16
1.3.3.2 CLR LIGANDS DERIVED FROM THE MYCOBACTERIAL CELL WALL 17
1.4 VACCINES AND AOJUVANTS 17
1.4.1 IMMUNITY AND VACCINATION AGAINST MTB 18
1.4.2 ADJUVANTS 21
I
HTTP://D-NB.INFO/1042265623
IMAGE 2
TABLE OF CONTENT
1.4.3 CATIONIC ADJUVANT FORMULATION CAFOI 22
2. MATERIAL FT METHODS 25
2.1 MATERIAL 25
2.1.1 CHEMICALS AND REAGENTS 25
2.1.2 KITS AND ENZYMES 26
2.1.3 WESTERN BLOT ANTIBODIES AND PROTEIN LADDER 26
2.1.3.1 WESTERN BLOT ANTIBODIES 26
2.1.4 PRIMERS AND PROBES 27
2.1.5 PLASMIDS 28
2.1.6 BUFFERS AND VACCINE FORMULATIONS 29
2.1.6.1 GRIESS ASSAY 29
2.1.6.2 EUSA 29
2.1.6.3 TRANSFECTION OF PHOENIX CELLS 29
2.1.6.4 FUSION PROTEIN BINDING ASSAY 29
2.1.6.5 SDS PAGE & WESTERN BLOT 29
2.1.6.6 CELL CULTURE 31
2.1.6.7 IMMUNIZATION 31
2.1.7 BACTERIA AND MEDIA 31
2.1.8 CELL CULTURE MEOIA, SUPPLEMENTS & STIMULATORY AGENTS 31
2.1.8.1 MEDIA 31
2.1.8.2 SUPPLEMENTS 31
2.1.8.3 COMPLETE DMEM (CDMEM) AND COMPLETE RPMI (CRPMI) 32
2.1.8.4 STIMULATORY AGENTS 32
2.1.9 CONSUMABLE ITEMS 32
2.1.10 LABORATORY EQUIPMENT 34
2.1.11 SOFTWARE 35
2.2 METHODS 36
2.2.1 MOLECULAR BIOLOGY 36
2.2.1.1 RNA ISOLATION 36
2.2.1.2 DETERMINATION OF NUCLEIC ACID CONCENTRATION & PURITY 36
2.2.1.3 CDNA SYNTHESIS 36
2.2.1.4 QUANTITATIVE REAL-TIME PCR & CALCULATION OF FOLD CHANGES 36
2.2.1.5 MICROARRAY ANALYSIS 38
2.2.1.6 POLYMERASE CHAIN REACTION 38
II
IMAGE 3
TABLE OF CONTENT
2.2.1.7 PREPARATION OF PLASMID DNA BY MINI OR MAXI PREP 38
2.2.2 DETECTION OF NITRIC OXIDE (GRIESS ASSAY) 38
2.2.3 IMMUNOASSAYS 38
2.2.3.1 ELISA 38
2.2.3.2 FUSION PROTEIN BINDING ASSAY 39
2.2.4 BIOCHEMICAL ANALYSIS 39
2.2.4.1 SDS-PAGE 39
2.2.4.2 WESTERN BLOT 39
2.2.4.3 PNGASE DIGEST 39
2.2.5 CELLCULTURE 40
2.2.5.1 CELL LINES 40
2.2.5.2 TRANSFECTION OF PHOENIX CELLS 40
2.2.5.3 L-CELL CONDITIONED MEDIUM (LCCM) 40
2.2.5.4 PREPARATION OF GM-CSF CONTAINING MEDIA 41
2.2.5.5 MACROPHAGE DIFFERENTIATION 41
2.2.5.6 DC DIFFERENTIATION 41
2.2.5.7 RETROVIRAL TRANSDUCTION OF DCS 41
2.2.5.8 PLATE COATING FOR MACROPHAGE & DC STIMULATION 42
2.2.5.9 STIMULATION WITH SOLUBLE STIMULI 43
2.2.5.10 PREPARATION OF BCG CULTURES FOR MACROPHAGE STIMULATION 43
2.2.5.11 ANTIGEN-SPECIFIC RESTIMULATION OF LYMPH NODE CELLS 44
2.2.6 CELL VIABILITY ASSAYS 44
2.2.6.1 MTT-ASSAY 44
2.2.6.2 LDH-ASSAY 44
2.2.7 MICE 44
2.2.7.1 STRAINS 44
2.2.7.2 IMMUNIZATION PROTOCOL 45
2.2.8 STATISTICAL EVALUATION 45
2.2.9 PROMOTER ANALYSIS 45
3. AIM OF THE STUDY 46
RESULTS 48
4.1 IDENTIFICATION OF THE PATTERN RECOGNITION RECEPTOR FOR THE
MYCOBACTERIAL CORD FACTOR TDM AND
ITS SYNTHETIC ANALOGUE TDB 48
III
IMAGE 4
TABLE OF CONTENT
4.1.1 RECOGNITION OF TDB AND TDM REQUIRES MLNCLE AND FCRGAMMA-CHAIN 48
4.1.1.1 RECOGNITION OF TDB AND TDM REQUIRES FCRGAMMA-CHAIN 48
4.1.1.2 IMMUNIZATION WITH CAFDL FAILS IN FCRGAMMA-/- MICE 49
4.1.1.3 MINDE EXPRESSION IS INDUCED IN MACROPHAGES UPON GLYCOLIPID
STIMULATION AND RECOMBINANT
MINDE BINDS TDB AND TDM 50
4.1.1.4 RECOGNITION OF TDB AND TDM BY MURINE MACROPHAGES DEPENDS ON
MINDE 52
4.1.1.5 EFFECTIVE IMMUNIZATION WITH CAFOL REQUIRES THE RECEPTOR MOLECULE
MINDE 56
4.1.2 MACROPHAGE ACTIVATION IN RESPONSE TO LIVING MYCOBACTERIA IS
IMPAIRED IN MINCIE-/- AND FCRG -/- --
58
4.1.2.1 RECOGNITION OF BCG IS PARTIALLY DEPENDENT ON FCRGAMMA-CHAIN 58
4.1.2.2 BCG-INDUCED MACROPHAGE ACTIVATION IS REDUCED IN MINDE-DEFICIENT
CELLS 59
4.1.2.3 PHAGOCYTOSIS OF BCG IS NOT IMPAIRED IN MINDE-DEFICIENT
MACROPHAGES 60
4.2 RESPONSIVENESS OF MACROPHAGES TO TDB STIMULATION DEPENDS ON MINCLE
EXPRESSION LEVELS,
C/EBPBETA AND HIFIALPHA 63
4.2.1 MYD88 REGULATES MINCLE EXPRESSION UPON TLR LIGATION AND THEREBY
INFLUENCES TDB-MEOIATED
MACROPHAGE ACTIVATION 63
4.2.1.1 MINCLE IS A GLYCOSYLATED PROTEIN STRONGLY INDUCED BY TDB AND LPS
63
4.2.1.2 LPS- OR CPG-INDUCED MINDE EXPRESSION IS IMPAIRED IN MYD88-/- 64
4.2.1.3 INCREASED MINCLE EXPRESSION CORRELATES WITH ENHANCED MACROPHAGE
RESPONSIVENESS TO TDB.
65
4.2.2 EXPRESSION PATTERNS AND REQUIREMENT OF INDUCIBLE TRANSCRIPTION
FACTORS 69
4.2.2.1 GLOBAL ANALYSIS OF TF EXPRESSION 70
4.2.2.2 EGRS ARE INDUCED EARLY AND INDEPENDENT OF NOVEL PROTEIN
BIOSYNTHESIS 71
4.2.2.3 HIFLA EXPRESSION IS REQUIRED FOR TDB-INDUCED NOS2 EXPRESSION AND
NO PRODUCTION 73
4.2.2.4 C/EBPBETA REGULATES MINCLE EXPRESSION AND IS REQUIRED FOR
TDB-INDUCED TARGET GENE
EXPRESSION 76
4.2.2.5 C/EBPBETA IS PARTIALLY REQUIRED FOR CURDLAN-INDUCED ACTIVATION
82
5. DISCUSSION 86
5.1 MINCLE EXPRESSION IS INDUCED UPON STIMULATION 86
5.2 MINCIE RECOGNIZES TDM AND TDB 89
5.3 MINCLE SIGNALING AND TRANSCRIPTION FACTOR ACTIVATION 91
5.4 TDB AS ADJUVANT 96
5.5 ROLE OF MINCLE IN EXPERIMENTAL TB INFECTION 101
IV
IMAGE 5
TABLE OF CONTENT
REFERENCES 106
DANKSAGUNG I
LIST OF PUBLICATIONS ILL
V
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| indexdate | 2024-07-10T00:42:21Z |
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| language | German English |
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| physical | XIX, 118, III S. Ill., graph. Darst. |
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| spelling | Schoenen, Hanne Gunda Verfasser aut Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion vorgelegt von Hanne Gunda Schoenen Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor 2013 XIX, 118, III S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2013 Mit einer Zssfassung in dt. Sprache (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026193042&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schoenen, Hanne Gunda Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |
| subject_GND | (DE-588)4113937-9 |
| title | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |
| title_alt | Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor |
| title_auth | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |
| title_exact_search | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |
| title_full | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion vorgelegt von Hanne Gunda Schoenen |
| title_fullStr | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion vorgelegt von Hanne Gunda Schoenen |
| title_full_unstemmed | Mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion vorgelegt von Hanne Gunda Schoenen |
| title_short | Mycobacterial cord factor mediated activation of murine macrophages |
| title_sort | mycobacterial cord factor mediated activation of murine macrophages analysis of the receptor and down stream signaling pathway aktivierung von mausmakrophagen durch den mykobakteriellen cord faktor untersuchung des rezeptors und der intrazellularen signaltransduktion |
| title_sub | analysis of the receptor and down-stream signaling pathway = Aktivierung von Mausmakrophagen durch den mykobakteriellen Cord-Faktor : Untersuchung des Rezeptors und der intrazellulären Signaltransduktion |
| topic_facet | Hochschulschrift |
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