Regulation of dendritic cell activation by apoptotic cell-derived annexin A1:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. Sprache |
| Physical Description: | XII, 130 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 |c presented by Veronika Jahndel |
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Record in the Search Index
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| adam_text | CONTENTS
DANKSAGUNG I
ABSTRACT V
ZUSAMMENFASSUNG VII
1 INTRODUCTION 1
1.1 THE VERTEBRATE IMMUNE SYSTEM 1
1.1.1 INNATE IMMUNITY 1
1.1.2 ADAPTIVE IMMUNITY 3
1.1.3 DC PRIME ADAPTIVE IMMUNITY 3
1.1.4 TOLERANCE TO SELF 5
1.2 DYING CELLS INFLUENCE THE IMMUNE SYSTEM 9
1.2.1 CLEARANCE OF DEAD CELLS BY PHAGOCYTES 10
1.2.2 TOLEROGENIC CELL DEATH 13
1.2.3 IMMUNOGENIC CELL DEATH 17
1.3 TOLL-LIKE RECEPTORS 18
1.3.1 TLR SIGNALLING 20
1.3.2 NEGATIVE REGULATION OF TLR SIGNALLING 23
1.3.3 REGULATION OF TLR SIGNALLING BY HETEROLOGOUS RECEPTORS 26
1.4 ANNEXIN A1 28
1.4.1 STRUCTURE OF ANXAL 29
1.4.2 ANXAL EXTERNALISATION AND PROTEOLYTIC CLEAVAGE 30
1.4.3 IMMUNOLOGICAL FUNCTIONS OF ANXAL 31
1.4.4 RECEPTORS FOR ANXAL 32
1.5 AIM OF THE STUDY 35
2 MATERIALS 37
2.1 CHEMICALS, REAGENTS AND CONSUMABLES 37
2.1.1 CONSUMABLES 37
2.1.2 COMMERCIAL KITS AND AUXILIARY EQUIPMENT 38
2.1.3 REAGENTS 38
IX
HTTP://D-NB.INFO/1044930616
CONTENTS
2.2 BUFFERS AND SOLUTIONS 39
2.3 CULTURE MEDIA 42
2.3.1 MEDIA AND SUPPLEMENTS FOR EUKARYOTIC CELL CULTURE 42
2.3.2 MEDIA FOR BACTERIA 42
2.4 BIOLOGICAL MATERIAL 43
2.4.1 EUKARYOTIC CELL LINE 43
2.4.2 BACTERIAL STRAIN 43
2.5 MATERIALS FOR MOLECULAR BIOLOGY 43
2.5.1 PRIMERS FOR QUANTITATIVE REAL-TIME PCR 43
2.5.2 VECTOR 44
2.6 ANTIBODIES 44
2.7 INSTRUMENTS 47
2.8 SOFTWARE 48
3 METHODS 49
3.1 METHODS IN MAMMALIAN CELL CULTURE 49
3.1.1 GENERAL CULTURE CONDITIONS 49
3.1.2 CULTURE OF JE6.1 JURKAT T CELLS 49
3.1.3 THAWING AND FREEZING OF JE6.1 JURKAT T CELLS 49
3.1.4 DETERMINATION OF CELL DENSITY AND VIABILITY 50
3.1.5 MONOCYTE ISOLATION FROM BLOOD AND DC DIFFERENTIATION 50
3.2 METHODS IN MOLECULAR BIOLOGY 51
3.2.1 TRANSFORMATION OF KCM-COMPETENT BACTERIA 51
3.2.2 ISOLATION OF TOTAL CELLULAR RNA 51
3.2.3 REVERSE TRANSCRIPTION OF RNA INTO CDNA 52
3.2.4 QUANTITATIVE REAL-TIME PCR 52
3.3 CELL BIOLOGICAL METHODS 54
3.3.1 APOPTOSIS INDUCTION AND ASSESSMENT 54
3.3.2 COCULTURE EXPERIMENTS OF DC WITH APOPTOTIC CELLS 54
3.3.3 EXPERIMENTS WITH DC USING RECOMBINANT ANXAL 55
3.4 BIOCHEMICAL AND IMMUNOLOGICAL METHODS 55
3.4.1 ANXAL EXPRESSION AND PURIFICATION OF ANXAL FROM E. COLI .... 55
3.4.2 CELL LYSIS AND PROTEIN QUANTIFICATION 56
3.4.3 PREPARATION OF NUCLEAR AND CYTOSOLIC CELL EXTRACTS 57
3.4.4 SDS-POLYACRYLAMIDE GEL ELECTROPHORESIS 57
3.4.5 SILVER STAINING OF SDS-POLYACRYLAMIDE GELS 58
X
CONTENTS
3.4.6 WESTERN BLOTTING 58
3.4.7 LIMULUS AMEBOCYTE LYSATE ASSAY 59
3.4.8 ENZYME-LINKED IMMUNOSORBENT ASSAY 59
3.4.9 IMMUNOFLUORESCENT STAINING FOR FLOW CYTOMETRY 60
3.5 STATISTICAL DATA ANALYSIS 61
4 RESULTS 63
4.1 ANXAL SUPPRESSES CYTOKINE SECRETION FROM HUMAN DC 63
4.1.1 ANXAL SUPPRESSES TLR-INDUCED CYTOKINE SECRETION 63
4.1.2 ANXAL REQUIRES PRE-INCUBATION FOR SUPPRESSION 65
4.1.3 ANXAL DOES NOT AFFECT CELL VIABILITY 66
4.1.4 ANXAL DOES NOT DOWNREGULATE TLR EXPRESSION 67
4.1.5 ANXAL DOES NOT INDUCE TNF CROSS-TOLERANCE 68
4.2 ANXAL INTERFERES WITH CYTOKINE TRANSCRIPTION 69
4.2.1 ANXAL REDUCES CYTOKINE TRANSCRIPT LEVELS 69
4.2.2 ANXAL DOES NOT ALTER TNF TRANSCRIPT STABILITY 71
4.3 ANXAL ATTENUATES TOLL-LIKE RECEPTOR SIGNALLING 72
4.3.1 ANXAL IMPAIRS ACTIVATION OF IKKA/3 AND THE MAPK CASCADE . . 72
4.3.2 ANXAL IMPAIRS SIGNALLING TO NF-KB BY TARGETING TAK1 74
4.4 ANXAL-TREATED DC SHOW FEATURES OF TOLEROGENICITY 77
4.4.1 ANXAL-TREATED DC SHOW AN IMMATURE PHENOTYPE 77
4.4.2 ANXAL-TREATED DC UPREGULATE TOLEROGENIC MOLECULES AND CHANGE
THEIR CHEMOKINE RECEPTOR EXPRESSION PATTERN 78
4.4.3 ANXAL ACTIVATES THE TRANSCRIPTION FACTOR STAT3 VIA JAK 81
4.4.4 ANXAL DOES NOT ACTIVATE AKT OR MAPK 83
4.4.5 ACTIVATED STAT3 MEDIATES UPREGULATION OF PD-L1 BUT IS DISPENS
ABLE FOR CYTOKINE SUPPRESSION BY ANXAL 84
5 DISCUSSION 87
5.1 CYTOKINE PROFILE OF ANXAL-TREATED DC 87
5.2 SUPPRESSION OF TLR SIGNALLING BY ANXAL 89
5.3 TOLEROGENIC POTENTIAL OF ANXAL-TREATED DC 93
5.4 ACTIVATION OF STAT3 BY ANXAL 96
5.5 PUTATIVE RECEPTORS FOR ANXAL ON DC 98
5.6 MODEL OF ANXAL FUNCTION 99
BIBLIOGRAPHY 103
XI
CONTENTS
APPENDIX 121
ABBREVIATIONS 121
ABBREVIATED PROTEINS 125
LIST OF FIGURES 129
LIST OF PUBLICATIONS 130
DECLARATION 130
XII
|
| any_adam_object | 1 |
| author | Jahndel, Veronika |
| author_GND | (DE-588)1044930403 |
| author_facet | Jahndel, Veronika |
| author_role | aut |
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| bvnumber | BV041476421 |
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| dewey-full | 571.96 |
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| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.96 |
| dewey-search | 571.96 |
| dewey-sort | 3571.96 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-10T00:57:39Z |
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| language | English |
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| physical | XII, 130 S. Ill., graph. Darst. |
| publishDate | 2013 |
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| spelling | Jahndel, Veronika Verfasser (DE-588)1044930403 aut Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 presented by Veronika Jahndel 2013 XII, 130 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache Heidelberg, Univ., Diss., 2013 Annexin I (DE-588)4548542-2 gnd rswk-swf Dendritische Zelle (DE-588)4313846-9 gnd rswk-swf Regulation (DE-588)4049075-0 gnd rswk-swf Apoptosis (DE-588)4291096-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dendritische Zelle (DE-588)4313846-9 s Regulation (DE-588)4049075-0 s Annexin I (DE-588)4548542-2 s Apoptosis (DE-588)4291096-1 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026922469&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Jahndel, Veronika Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 Annexin I (DE-588)4548542-2 gnd Dendritische Zelle (DE-588)4313846-9 gnd Regulation (DE-588)4049075-0 gnd Apoptosis (DE-588)4291096-1 gnd |
| subject_GND | (DE-588)4548542-2 (DE-588)4313846-9 (DE-588)4049075-0 (DE-588)4291096-1 (DE-588)4113937-9 |
| title | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 |
| title_auth | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 |
| title_exact_search | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 |
| title_full | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 presented by Veronika Jahndel |
| title_fullStr | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 presented by Veronika Jahndel |
| title_full_unstemmed | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 presented by Veronika Jahndel |
| title_short | Regulation of dendritic cell activation by apoptotic cell-derived annexin A1 |
| title_sort | regulation of dendritic cell activation by apoptotic cell derived annexin a1 |
| topic | Annexin I (DE-588)4548542-2 gnd Dendritische Zelle (DE-588)4313846-9 gnd Regulation (DE-588)4049075-0 gnd Apoptosis (DE-588)4291096-1 gnd |
| topic_facet | Annexin I Dendritische Zelle Regulation Apoptosis Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026922469&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT jahndelveronika regulationofdendriticcellactivationbyapoptoticcellderivedannexina1 |