The phenotypic and functional analysis of multinucleated giant cells:
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
München
Verl. Dr. Hut
2014
|
| Edition: | 1. Aufl. |
| Series: | Medizin
|
| Subjects: | |
| Online Access: | Inhaltstext Inhaltsverzeichnis |
| Physical Description: | XVI, 158 S. Ill., graph. Darst. 210 mm x 148 mm, 276 g |
| ISBN: | 9783843915359 3843915350 |
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MARC
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| 100 | 1 | |a Milde, Ronny |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a The phenotypic and functional analysis of multinucleated giant cells |c Ronny Milde |
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| 264 | 1 | |a München |b Verl. Dr. Hut |c 2014 | |
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Record in the Search Index
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VN
TABLE OF CONTENTS
LIST OF FIGURES XI
LIST OF TABLES XV
1 INTRODUCTION . 1
1.1 MACROPHAGES AS PART OF THE INNATE IMMUNE SYSTEM 1
1.2 THE FUNCTION OF MACROPHAGES 5
1.2.1 PHAGOCYTOSIS 5
1.2.1.1 THE PROCESS OF PHAGOCYTOSIS 5
1.2.1.2 PHAGOCYTIC RECEPTORS 6
1.2.2 ANTIGEN PRESENTATION 10
1.2.3 SECRETION OF INFLAMMATORY MOLECULES 10
1.2.4 TISSUE HOMEOSTASIS 10
1.3 MACROPHAGE ACTIVATION 11
1.4 MACROPHAGE FUSION AND MULTINUCLEATED GIANT CELLS 15
1.4.1 MULTINUCLEATED GIANT CELLS 15
1.4.2 THE MECHANISM OF MACROPHAGE FUSION 17
1.4.3 THE FUNCTION OF FUSED MACROPHAGES 18
1.4.3.1 OSTEOCLASTS 18
1.4.3.2 MULTINUCLEATED GIANT CELLS 19
1.5 AIM OF THE WORK 24
2 MATERIALS AND METHODS 25
2.1 MATERIALS 25
2.1.1 BACTERIAL STRAINS 25
2.1.2 MICE 25
2.1.3 CELL LINES 25
2.1.4 HUMAN TUBERCULOSIS SAMPLES 25
2.1.5 CHEMICALS/ REAGENTS 26
2.1.6 OLIGODEOXYNUCLEOTIDES 28
2.1.6.1 OLIGODEOXYNUCLEOTIDES FOR SYBR GREEN QPCR METHOD 28
2.1.6.2 OLIGODEOXYNUCLEOTIDES FOR TAQMAN QPCR METHOD 29
HTTP://D-NB.INFO/1050732022
VNI
2.1.7 ANTIBODIES AND ENZYMES 31
2.1.7.1 ANTIBODIES FOR FLOW CYTOMETRY 31
2.1.7.2 BLOCKING ANTIBODIES 31
2.1.7.3 ANTIBODIES USED FOR ANALYSIS
OF PHAGOCYTOSIS 32
2.1.7.4 ANTIBODIES USED FOR IMMUNOHISTOCHEMISTRY 32
2.1.7.5 ANTIBODIES USED IN WESTERN BLOT ANALYSES 32
2.1.7.6 ENZYMES 33
2.1.8 ANTIBIOTIC AGENTS 33
2.1.9 CYTOKINES 33
2.1.10 BUFFERS, SOLUTIONS AND MEDIA 33
2.1.11 INHIBITORS 38
2.1.12 STANDARDS AND KITS 38
2.1.13 EQUIPMENT 38
2.1.14 OTHER MATERIALS 39
2.2 METHODS 40
2.2.1 SCHISTOSOMA MCMSONI INFECTION OF MICE 40
2.2.2 ISOLATION OF PRIMARY MACROPHAGES 40
2.2.3 GENERATION OF MULTINUCLEATED GIANT CELLS 40
2.2.4 SEPARATION OF MONONUCLEATED MACROPHAGES
AND MGCS BY SIEVING 41
2.2.5 FLOW CYTOMETRY 42
2.2.6 ADHESION ASSAY AND CRYSTAL VIOLET STAINING 42
2.2.7 LYSIS OF CELLS AND MEASUREMENT OF PROTEIN CONCENTRATION 43
2.2.8 WESTERN BLOT ANALYSIS 43
2.2.9 NO ASSAY 44
2.2.10 ARGINASE ASSAY 44
2.2.11 ELISAANDCBA 45
2.2.12 ZYMOGRAPHY 45
2.2.13 MICROARRAY ANALYSIS 46
2.2.14 PROTEOME ANALYSIS 47
2.2.15 OPSONIZATION OF SHEEP RED
BLOOD CELLS 48
2.2.16 COATING OF POLYSTYRENE BEADS 48
2.2.17 FITC-LABELING OF ZYMOSAN 49
IX
2.2.18 CULTIVATION OF LISTERIA MONOCYTOGENES 49
2.2.19 PHAGOCYTOSIS ASSAY 49
2.2.20 HEMACOLOR STAINING 50
2.2.21 IMMUNOSTAINING OF CULTURED
CELLS SI
2.2.22 GELATIN DEGRADATION ASSAY 51
2.2.23 BONE DEGRADATION ASSAY 52
2.2.24 SNAP-FREEZING OF ORGAN TISSUE SAMPLES (* FRESH FROZEN MATERIAL)
52
2.2.25 FIXATION AND PARAFFIN EMBEDDING
OF ORGAN TISSUE SAMPLES (** FFPE-MATERIAL).
52
2.2.26 CRYOSECTIONING 53
2.2.27 SECTIONING OF FFPE TISSUE SAMPLES 53
2.2.28 HEMATOXYLIN AND EOSIN STAIN (HE) 53
2.2.29 MASSON'S TRICHROME STAIN 53
2.2.30 ZIEHL-NEELSON STAIN 54
2.2.31 IMMUNOSTAINING OF TISSUE SECTIONS 54
2.2.32 LASER DISSECTION MICROSCOPY
(LDM) 55
2.2.33 RNA PREPARATION 56
2.2.34 REVERSE TRANSCRIPTION AND REAL
TIME PCR
(QPCR) 57
2.2.35 COMPUTATIONAL AND STATISTICAL ANALYSIS 57
3
RESULTS.
59
3.1 ANALYSIS OF MGCS
IN VITRO 59
3.1.1 ESTABLISHMENT OF A METHOD FOR THE SEPARATION OF MACROPHAGES AND
MGCS 60
3.1.2 PHENOTYPIC ANALYSIS OF MGCS 66
3.1.2.1 MRNA EXPRESSION AND PROTEOME ANALYSIS 66
3.1.3 FUNCTIONAL ANALYSIS OF MGCS 73
3.1.3.1 PHAGOCYTIC CAPACITY OF MGCS 73
3.1.3.2 ADHESION OF MGCS
ON BACTERIOLOGICAL PLASTIC IS DEPENDENT
ON CD
18/CD11
C95
3.1.3.3 ARGINASE ACTIVITY
AND NO PRODUCTION ARE NORMAL IN MGCS 98
3.1.3.4 MGCS SHOW ALTERED LPS PHENOTYPE 100
3.1.3.5 MGCS SHOW AN INCREASED EXPRESSION
OF DEGRADATIVE MMPS 104
3.2 ANALYSIS OF MGCS
IN VIVO 108
3.2.1 MGCS IN HUMAN TUBERCULOSIS AND MURINE S. MANSONI INFECTION 108
3.2.2 PHENOTYPIC ANALYSIS 108
X
3.2.2.1 SEPARATION OF MACROPHAGES AND MGCS FROM TISSUE SAMPLES USING
LASER
DISSECTION MICROSCOPY (LDM) 112
3.2.2.2 MGCS SHOW A UNIQUE MRNA EXPRESSION WITH INCREASED EXPRESSION OF
CART,
CDLLC AND DC-STAMP 116
3.2.2.3 RNA EXPRESSION ARRAY
OF MGCS AND MACROPHAGES
DISSECTED BY LDM 119
4
DISCUSSION.*.
125
4.1 ESTABLISHMENT OF AN IN VITRO SEPARATION METHOD FOR THE ENRICHMENT OF
MGCS AND
MACROPHAGES FROM MIXED CULTURES 12S
4.2 MGCS SHOW A DISTINCT PHENOTYPE IN PHAGOCYTOSIS 126
4.3 MGCS SHOW AN INCREASE IN THE
SECRETION OF DEGRADATIVE
ENZYMES 132
4.4 MGCS SHOW AN ATTENUATED LPS RESPONSE 134
4.5 GENES REGULATED IN MGCS IN VITRO: FUNCTIONAL INDICATIONS 13S
4.6 MGCS IN TISSUE SECTIONS
OF TUBERCULOSIS AND SCHISTOSOMIASIS 138
4.7 TRANSCRIPTOME ANALYSIS OF TISSUE
MGCS 139
5
ABBREVIATIONS.
143
6 REFERENCES
.145 |
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| author | Milde, Ronny |
| author_facet | Milde, Ronny |
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| bvnumber | BV041947272 |
| classification_tum | BIO 200d |
| ctrlnum | (OCoLC)879878951 (DE-599)DNB1050732022 |
| discipline | Biologie Medizin |
| edition | 1. Aufl. |
| format | Thesis Book |
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| isbn | 9783843915359 3843915350 |
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| physical | XVI, 158 S. Ill., graph. Darst. 210 mm x 148 mm, 276 g |
| publishDate | 2014 |
| publishDateSearch | 2014 |
| publishDateSort | 2014 |
| publisher | Verl. Dr. Hut |
| record_format | marc |
| series2 | Medizin |
| spelling | Milde, Ronny Verfasser aut The phenotypic and functional analysis of multinucleated giant cells Ronny Milde 1. Aufl. 201405 München Verl. Dr. Hut 2014 XVI, 158 S. Ill., graph. Darst. 210 mm x 148 mm, 276 g txt rdacontent n rdamedia nc rdacarrier Medizin Zugl.: München, Techn. Univ., Diss., 2013 Phagozytose (DE-588)4129146-3 gnd rswk-swf Riesenzelle (DE-588)4304362-8 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Riesenzelle (DE-588)4304362-8 s Phagozytose (DE-588)4129146-3 s DE-604 X:MVB text/html http://deposit.dnb.de/cgi-bin/dokserv?id=4658237&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027390316&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Milde, Ronny The phenotypic and functional analysis of multinucleated giant cells Phagozytose (DE-588)4129146-3 gnd Riesenzelle (DE-588)4304362-8 gnd |
| subject_GND | (DE-588)4129146-3 (DE-588)4304362-8 (DE-588)4113937-9 |
| title | The phenotypic and functional analysis of multinucleated giant cells |
| title_auth | The phenotypic and functional analysis of multinucleated giant cells |
| title_exact_search | The phenotypic and functional analysis of multinucleated giant cells |
| title_full | The phenotypic and functional analysis of multinucleated giant cells Ronny Milde |
| title_fullStr | The phenotypic and functional analysis of multinucleated giant cells Ronny Milde |
| title_full_unstemmed | The phenotypic and functional analysis of multinucleated giant cells Ronny Milde |
| title_short | The phenotypic and functional analysis of multinucleated giant cells |
| title_sort | the phenotypic and functional analysis of multinucleated giant cells |
| topic | Phagozytose (DE-588)4129146-3 gnd Riesenzelle (DE-588)4304362-8 gnd |
| topic_facet | Phagozytose Riesenzelle Hochschulschrift |
| url | http://deposit.dnb.de/cgi-bin/dokserv?id=4658237&prov=M&dok_var=1&dok_ext=htm http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027390316&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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