Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
Berlin
2012
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | X, 131 S. Ill., graph. Darst. |
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MARC
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| 245 | 1 | 0 | |a Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |c von Kathleen D. Metzler |
| 264 | 1 | |a Berlin |c 2012 | |
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| 502 | |a Berlin, Humboldt-Univ., Diss., 2012 | ||
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|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS III
T A B L E OF CONTENTS
CHAPTER 1: INTRODUCTION 1
1.1 NEUTROPHILS 1
1.1.1 MORPHOLOGY 1
1.1.2 CHALLENGES O F STUDYING NEUTROPHILS 2
1.2 NEUTROPHIL GRANULES 2
1.2.1 GRANULE TYPES 2
1.2.2 SERGLYCIN 4
1.3 NEUTROPHIL ARSENAL 5
1.3.1 REACTIVE OXYGEN SPECIES AND REACTIVE NITROGEN SPECIES 5
1.3.2 MYELOPEROXIDASE 9
1.3.3 NEUTROPHIL SERINE PROTEASES 14
1.4 NEUTROPHIL ANTIMICROBIAL ACTIVITIES 19
1.4.1 RECRUITMENT O F NEUTROPHILS TO INFECTED TISSUES 19
1.4.2 NEUTROPHIL ANTIMICROBIAL MECHANISMS 20
1.5 NEUTROPHIL EXTRACELLULAR TRAPS (NETS) 22
1.5.1 NETS AND PATHOGENS 22
1.5.2 DETRIMENTAL EFFECTS O F NETS 24
1.5.3 MECHANISM O F N E T FORMATION 25
1.6 AIM O F THE STUDY 30
CHAPTER 2 : MATERIALS AND M E T H O D S 3 1
2.1 BUFFERS AND SOLUTIONS 31
2.2 PHARMACOLOGICAL INHIBITORS 33
2.3 CELL ISOLATION AND CELL CULTURE 34
2.3.1 DONOR CONSENT 34
2.3.2 NEUTROPHIL ISOLATION 34
HTTP://D-NB.INFO/1033874329
IMAGE 2
I V TABLE OF CONTENTS
2.3.3 PBMC ISOLATION 34
2.3.4 CULTURE O F HELA CELLS 35
2.3.5 CULTURE O F HL-60 CELLS 35
2.4 STIMULATION O F NEUTROPHILS TO MAKE NETS 35
2.4.1 STIMULATION WITH PMA 35
2.4.2 STIMULATION WITH G O 35
2.4.3 STIMULATION WITH C. ALBICANS 36
2.4.4 STIMULATION WITH CHLORAMINES 36
2.4.5 TRANSWELL EXPERIMENTS 36
2.4.6 CELL CO-INCUBATION EXPERIMENTS 36
2.5 MICROBIOLOGY METHODS 36
2.5.1 CANDIDA ALBICANS 36
2.5.2 SHIGELLA FLEXNERI 37
2.6 MICROSCOPY METHODS 38
2.6.1 IMMUNOFLUORESCENCE STAINING 38
2.6.2 VISUALIZATION AND QUANTIFICATION O F N E T FORMATION 39
2.6.3 EPIFLUORESCENCE MICROSCOPY 39
2.6.4 CONFOCAL MICROSCOPY 40
2.6.5 SCANNING ELECTRON MICROSCOPY 40
2.7 ENZYMATIC ACTIVITY METHODS 41
2.7.1 LUMINOL ASSAY FOR ROS PRODUCTION 41
2.7.2 CYTOTOXICITY ASSAY 41
2.7.3 PROTEASE ACTIVITY ASSAYS 41
2.7.4 MYELOPEROXIDASE ACTIVITY ASSAY 42
2.8 BIOCHEMICAL METHODS 42
J
2.8.1 BASIC BIOCHEMICAL METHODS 42
2.8.2 PROTEIN PURIFICATION 44
2.8.3 IMMUNOPRECIPITATION EXPERIMENTS 44
2.8.4 PROTEIN IDENTIFICATION BY MASS SPECTROMETRY 45
IMAGE 3
TABLE OF CONTENTS V
2.8.5 REACTIVE OXYGEN SPECIES 45
2.8.6 SUBCELLULAR FRACTIONATION 46
2.8.7 MEMBRANE PERMEABILIZATION EXPERIMENTS USING SYNTHETIC LIPOSOMES 4
8
2.8.8 GRANULE PERMEABILIZATION EXPERIMENTS 4 8
2.9 ANIMAL EXPERIMENTS 49
2.9.1 APPROVAL O F ANIMAL EXPERIMENTS 49
2.9.2 MOUSE LUNG INFECTIONS 49
2.9.3 ISOLATION O F MOUSE PERITONEAL CELLS 49
2.10 STATISTICAL ANALYSIS 49
CHAPTER 3 : MYELOPEROXIDASE IS REQUIRED FOR N E T FORMATION 5 0
3.1 CHARACTERIZATION O F MPO-DEFICIENT DONORS 50
3.2 M P O IS REQUIRED FOR N E T FORMATION 52
3.3 M P O ACTS CELL AUTONOMOUSLY TO PROMOTE N E T FORMATION 54
3.4 NET-DEPENDENT INHIBITION O F C. ALBICANS GROWTH IS COMPROMISED IN
MPO-DEFICIENT NEUTROPHILS 59
CHAPTER 4 : NEUTROPHIL ELASTASE AND MYELOPEROXIDASE REGULATE N E T
FORMATION 6 2
4.1 NEUTROPHIL EXTRACTS PROMOTE CHROMATIN DECONDENSATION 62
4.2 N E ACTIVITY IS REQUIRED FOR CHROMATIN DECONDENSATION 62
4.3 N E DEGRADES HISTONES, PROMOTING NUCLEAR DECONDENSATION 64
4.4 N E ACTIVITY IS REQUIRED FOR N E T FORMATION 65
4.5 N E CLEAVES HISTONES DURING N E T FORMATION 68
4.6 N E TRANSLOCATES TO THE NUCLEUS DURING N E T FORMATION 70
4.7 -NE IS REQUIRED FOR N E T FORMATION IN VIVO 72
CHAPTER 5 : T H E AZUROSOME PERMEABILIZES GRANULE MEMBRANES,
PROMOTING N E T FORMATION 7 8
5.1 REACTIVE OXYGEN SPECIES ARE REQUIRED F O R N E RELEASE FROM
AZUROPHILIC GRANULES 78
IMAGE 4
V I
TABLE OF CONTENTS
5.2 CHARACTERIZATION O F THE AZUROSOME: A COMPLEX O F AZUROPHILIC
GRANULE PROTEINS ...84
5.3 THE AZUROSOME PERMEABILIZES MEMBRANES 87
CHAPTER 6 : M P 0 / H 2 0 2 SYSTEM REGULATES NEUTROPHIL PROTEASES 9 3
6.1 MPO/H2O2 SYSTEM ACTIVATES PROTEASES IN NEUTROPHIL GRANULES 93
6.2 MPO/H2O2 SYSTEM REGULATES PURIFIED GRANULE PROTEASES AGAINST CERTAIN
SUBSTRATES..96
6.3 AZUROSOME PROTEASES ARE REGULATED BY MPO/H2O2 SYSTEM 97
CHAPTER 7 : DISCUSSION 1 0 1
7.1 M P O AND N E T FORMATION 101
7.1.1 M P O IS REQUIRED FOR N E T FORMATION 101
7.1.2 M P O ACTS CELL-AUTONOMOUSLY DURING N E T FORMATION 102
7.1.3 MULTIPLE ROLES O F M P O IN N E T FORMATION 102
7.1.4 MPO, N E T S AND IMMUNITY 103
7.2 CHROMATIN DECONDENSATION DURING N E T FORMATION 106
7.2.1 NEUTROPHIL SERINE PROTEASES IN N E T FORMATION 106
7.2.2 HISTONE MODIFICATIONS IN N E T FORMATION 107
7.2.3 NEUTROPHILS CAN UNDERGO NETOSIS DURING PHAGOCYTOSIS 107
7.3 HOW DO ROS AND N E REGULATE N E T FORMATION? 108
7.3.1 TRANSLOCATION O F GRANULE CONTENTS TO THE NUCLEUS 108
7.3.2 AZUROSOME-MEDIATED MEMBRANE PERMEABILIZATION 109
7.3.3 ROS AND PROTEASE REGULATION 110
7.4 CONCLUSION 113
REFERENCES 1 1 4
APPENDIX 1 2 8
CURRICULUM VITAE 128
EDUCATION: 128
IMAGE 5
TABLE OF CONTENTS V I I
RESEARCH EXPERIENCE: 128
TEACHING: 128
PUBLICATIONS 129
PUBLICATIONS IN JOURNALS: 129
PUBLICATIONS AT MEETINGS: 129
SELBSTANDIGKEITSERKLARUNG 130
ACKNOWLEDGEMENTS 1 3 1
|
| any_adam_object | 1 |
| author | Metzler, Kathleen D. 1983- |
| author_GND | (DE-588)1030129533 |
| author_facet | Metzler, Kathleen D. 1983- |
| author_role | aut |
| author_sort | Metzler, Kathleen D. 1983- |
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| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
| dewey-search | 610 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| physical | X, 131 S. Ill., graph. Darst. |
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| spelling | Metzler, Kathleen D. 1983- Verfasser (DE-588)1030129533 aut Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps von Kathleen D. Metzler Berlin 2012 X, 131 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Berlin, Humboldt-Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content Reproduziert als Metzler, Kathleen D. Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps Ketsch bei Mannheim : Mikroform Dissertation, 2012 2 Mikrofiches (DE-604)BV041075895 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025676885&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Metzler, Kathleen D. 1983- Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| subject_GND | (DE-588)4113937-9 |
| title | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| title_auth | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| title_exact_search | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| title_full | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps von Kathleen D. Metzler |
| title_fullStr | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps von Kathleen D. Metzler |
| title_full_unstemmed | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps von Kathleen D. Metzler |
| title_short | Myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| title_sort | myeloperoxidase and neutrophil elastase collaborate in the formation of neutrophil extracellular traps |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025676885&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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