Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen: = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | German |
| Published: |
Erlangen ; Nürnberg
[2018]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | IV, 164 Seiten Illustrationen, Diagramme 21 cm |
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MARC
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| 245 | 1 | 0 | |a Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen |b = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |c vorgelegt von Jennifer J. Lühr |
| 246 | 1 | 3 | |a Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| 246 | 1 | 1 | |a Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| 264 | 1 | |a Erlangen ; Nürnberg |c [2018] | |
| 300 | |a IV, 164 Seiten |b Illustrationen, Diagramme |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Friedrich-Alexander-Universität Erlangen-Nürnberg |d 2018 | ||
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-030900185 | ||
Record in the Search Index
| _version_ | 1804179463053770752 |
|---|---|
| adam_text | TABLE
OF
CONTENT
1
ZUSAMMENFASSUNG
................................................................................................................
1
2
SUMMARY
................................................................................................................................
2
3
INTRODUCTION
.............................................................................................................................
3
3.1
THE
IMMUNE
SYSTEM
.......................................................................................................
3
3.1.1
INNATE
IMMUNITY
........................................................................................................
3
3.1.2
ADAPTIVE
IMMUNITY
....................................................................................................
4
3.2
DENDRITIC
CELLS
..................................................................................................................
5
3.2.1
IDENTIFICATION
OF
DENDRITIC
CELLS
.................................................................................
5
3.2.2
DEVELOPMENT
OF
DENDRITIC
CELLS
................................................................................
5
3.2.3
ANTIGEN
UPTAKE,
ACTIVATION,
AND
PROCESSING
IN
DENDRITIC
CELLS
...............................
6
3.2.4
ACTIVATION
OF
T
CELLS
BY
DENDRITIC
CELLS
.....................................................................
9
3.2.5
FORMATION
OF
THE
IMMUNOLOGICAL
SYNAPSE
.............................................................
10
3.3.
C-TYPE
LECTIN
RECEPTORS
.................................................................................................
12
3.3.1
DEC205
..................................................................................................................
14
3.3.2
DCIR
.......................................................................................................................
14
3.3.3
MMR
.......................................................................................................................
15
3.3.4
DC-SIGN
.................................................................................................................
15
3.4
ANTIGEN
TARGETING
TO
DENDRITIC
CELLS
...............................................................................
15
3.4.1
MONOCYTE-DERIVED
DENDRITIC
CELLS
...........................................................................
16
3.4.2
DELIVERY
OF
ANTIGENS
TO
DENDRITIC
CELLS
BY
USAGE
OF
RECOMBINANT
ANTIBODIES
......
17
3.4.3
ANTIGEN
TARGETING
TO
DCS
HARNESSING
C-TYPE
LECTIN
RECEPTORS
..............................
18
3.5
BIOMEMBRANES
..............................................................................................................
19
4
AIMS
.....................................................................................................................................
21
5
RESULTS
..................................................................................................................................
22
5.1
EXPRESSION
OF
C-TYPE
LECTIN
RECEPTORS
ON
MONOCYTE-DERIVED
DENDRITIC
CELLS
.............
22
5.2
SPATIAL
LOCALIZATION
OF
CLRS
IN
THE
MODCS
MEMBRANE
..............................................
26
5.3
SEQUENCE
ALIGNMENT
OF
CLRS
......................................................................................
38
5.3.1
COMPARISON
OF
THE
INTRACELLULAR
DOMAINS
OF
DEC205,
DCIR,
MMR,
AND
DC-
SIGN
.................................................................................................................................
38
5.3.2
COMPARISON
OF
THE
TRANSMEMBRANE
REGIONS
OF
DEC205,
DCIR,
MMR,
AND
DC-
SIGN
.................................................................................................................................40
5.3.3
COMPARISON
OF
THE
HINGE
REGIONS
OF
DEC205,
DCIR,
MMR,
AND
DC-SIGN
...... 42
5.3.4
OVERALL
COMPARISON
OF
THE
STRUCTURAL
DOMAINS
OF
DEC205,
DCIR,
MMR,
AND
DC-
SIGN
.................................................................................................................................45
5.4
LIPID
ANALYSIS
OF
IMMATURE
AND
MATURE
MODCS
.......................................................... 46
5.4.1
IMMATURE
MODCS
DISPLAY
A
MORE
RIGID
MEMBRANE
THAN
MATURE
MODCS
...........46
5.4.2
CHOLESTEROL
ANALYSIS
OF
IMMATURE
AND
MATURE
MODCS
........................................47
5.4.3
LIPIDOMICS
ANALYSES
OF
IMMATURE
AND
MATURE
MODCS
........................................
48
5.4.3.1
LIPID
ANALYSIS
OF
SERUM
SAMPLES
....................................................................
49
5.4.3.2
LIPIDOMICS
ANALYSES
OF
IMMATURE
AND
MATURE
MODCS
.................................
52
5.4.4
LIPID
RAFT
ANALYSIS
IN
IMMATURE
AND
MATURE
MODCS
.............................................
66
5.5.
ANALYSIS
OF
PIP2
AND
DAG
IN
IMMATURE
AND
MATURE
MODCS
....................................
71
5.6
INFLUENCE
OF
F-ACTIN
ON
CLR
LOCALIZATION
.....................................................................
82
5.7
ANALYSIS
OF
ENERGY
METABOLISMS
IN
IMMATURE
AND
MATURE
MODCS
...........................
86
5.8
MIGRATION
OF
IMMATURE
AND
MATURE
MODCS
.................................................................
92
6
OUTLOOK
..................................................................................................................................
95
7
DISCUSSION
............................................................................................................................
97
7.1
INVESTIGATION
OF
LECTIN
RECEPTOR
EXPRESSION
ON
MONOCYTE-DERIVED
DCS
....................
97
7.2
SPATIAL
DISTRIBUTION
OF
CLRS
IN
THE
MEMBRANE
OF
MODCS
..........................................
99
7.2.1
IMMUNOFLUORESCENCE
ANALYSES
OF
CLRS
...............................................................
99
7.2.2
UPREGULATION
OF
DEC205
IN
MATURE
MODCS
........................................................
102
7.3
HOMOLOGY
STUDIES
OF MOTIF
SEQUENCES
OF
CLRS
.........................................................
102
7.4
LIPID
ANALYSIS
OF
IMMATURE
AND
MATURE
MODC
..........................................................
104
7.4.1
LIPID
PACKING
IMAGING
OF
IMMATURE
AND
MATURE
MODCS
...................................
104
7.4.2
LIPIDOMICS
ANALYSES
OF
IMMATURE
AND
MATURE
MODCS
......................................
105
7.4.3
LIPID
RAFT
ANALYSES
OF
IMMATURE
AND
MATURE
MODCS
..........................................
107
7.4.4
GENERAL
REMARKS
ABOUT
LIPID
ANALYSES
................................................................
108
7.5
TRANSIENT
EXPRESSION
OF
PIP2
AND
DAG
IN
MODCS
...................................................
109
7.6
F-ACTIN
IN
THE
MEMBRANE
OF
MODCS
...........................................................................
110
7.7
FUNCTIONAL
STUDIES
OF
MODCS
.....................................................................................
111
7.7.1
MODCS
ARE
DEPENDENT
ON
AEROBIC
GLYCOLYSIS
....................................................
111
7.7.2
MIGRATION
OF
IMMATURE
AND
MATURE
MODCS
.........................................................
111
8
MATERIAL
AND
METHODS
........................................................................................................
113
8.1
MATERIAL
..........................................................................................................................
113
8.1.1
CHEMICALS
.............................................................................................................
113
8.1.2
BUFFERS
AND
REAGENTS
............................................................................................
115
8.1.3
KITS
.........................................................................................................................
116
8.1.4
MOLECULAR
STANDARDS
FOR
GELS
................................................................................
116
8.1.5
ENZYMES
...............................................................................................................
116
8.1.6
CELL
LINES
...............................................................................
117
8.1.7
PLASMIDS
...............................................................................................................
117
8.1.8
PRIMER
...................................................................................................................
118
8.1.8.1
PRIMERS
FOR
CLONING
.........................................................................................
118
8.1.8.2
SEQUENCING
PRIMERS
......................................................................................
119
8.1.9
ANTIBODIES
.............................................................................................................
119
8.1.9.1
PRIMARY
ANTIBODIES
.........................................................................................
119
8.1.9.2
SECONDARY
ANTIBODIES
AND
STREPTAVIDIN
CONJUGATES,
AND
OTHERDYES
........
120
8.1.9.3
ISOTYPE
CONTROLS
.............................................................................................
120
8.1.10
ELECTRONIC
DEVICES
..............................................................................................
120
8.1.11
CENTRIFUGES
.........................................................................................................
121
8.1.12
MICROSCOPES
.......................................................................................................
121
8.1.13
CYTOMETERS
.........................................................................................................
122
8.1.14
SOFTWARE
..............................................................................................................
122
8.2
METHODS
....................................................................................................................
124
8.2.1
CLONING
OF
PLASMIDS
..............................................................................................
124
8.2.1.1
PCR
OF
DNA
FRAGMENTS
................................................................................
124
8.2.1.2
AGAROSE
GEL
ELECTROPHORESIS
AND
DNA
GEL
EXTRACTION
.................................
124
8.2.1.3
DNA
DIGESTION
WITH
RESTRICTION
ENZYMES
......................................................
125
8.2.1.4
LIGATION
...........................................................................................................
125
8.2.1.5
COMPETENT
BACTERIA
......................................................................................
126
8.2.1.6
TRANSFORMATION
IN
COMPETENT
BACTERIA
.........................................................
126
8.2.1.7
DNA
MINI
PREPARATION
...................................................................................
127
8.2.1.8
SEQUENCING
...................................................................................................
127
8.2.1.9
MAXI
PREPARATION
OF
DNA
..............................................................................
127
8.2.1.10
GLYCEROL
STOCK
OF
BACTERIA
...........................................................................
128
8.2.2
CELL
CULTURE
............................................................................................................
128
8.2.2.1
SPLITTING
OF
HEK293T
CELLS
...........................................................................
128
8.2.2.2
FREEZING
AND
THAWING
OF
HEK293T
CELLS
.....................................................
129
8.2.3
ANTIBODY
PRODUCTION
.............................................................................................
129
8.2.3.1
CALCIUM
PHOSPHATE
TRANSFECTION
OF
HEK293T
CELLS
....................................
129
8.2.3.2
AMMONIUM
SULFATE
PRECIPITATION
AND
DIALYSIS
..............................................
130
8.2.3.3
PROTEIN
G
BEADS
AND
ELUTION
OF
ANTIBODIES
..................................................
131
8.2.3
4
SDS-PAGE,
COOMASSIE
STAINING,
AND
DRYING
............................................
131
8.2.3.5
CASSETTE
DIALYSIS
AND
LPS
REMOVAL
.............................................................
132
8.2.3.6
PROTEIN
MEASUREMENT
...................................................................................
133
8.2.3
7
LPS
TESTING
....................................................................................................
133
8.2.3
8
WESTERN
BLOT
..................................................................................................
133
8.2.4
OVEREXPRESSION
OF
PIP2
AND
DAG
IN
CELL
CULTURE
...............................................
134
8.2.4.1
X-TREMEGENE
9
TRANSFECTION
.......................................................................
134
8.2.4
2
LINEARIZATION
AND
PURIFICATION
OF
VECTORS
FOR
RNA
TRANSFECTION
...................
134
8.2.4.3
MRNA
GENERATION
..........................................................................................
135
8.2.4
4
ELECTROPORATION
OF
MRNA
INTO
MODCS
..........................................................
135
8.2.5
GENERATION
OF
MONOCYTE-DERIVED
DENDRITIC
CELLS
.................................................
136
8.2.6
CELL
COUNTING
.........................................................................................................
137
8.2.7
IMMUNOFLUORESCENCE
STAINING
..............................................................................
137
8.2.7.1.
CO-STAINING
OF
CLRS
IN
THE
DC
MEMBRANE
.................................................
137
8.2.7.2
LIPID
RAFT
STAINING
...........................................................................................
139
8.2.7.3
TIRE
MICROSCOPY
...........................................................................................
139
8.2.8
FLOW
CYTOMETRY
ANALYSIS
OF
MODCS
.....................................................................
139
8.2.9
LIPIDOMIC
ANALYSIS
OF
MODCS
..............................................................................
141
8.2.10
CHOLESTEROL
ASSAY
..............................................................................................
142
8.2.11
AGILENT
XF
MITO
FUEL
FLEX
TEST
..........................................................................
142
8.2.12
MIGRATION
ASSAY
OF
IMMATURE
AND
MATURE
MODCS
............................................
143
9
REFERENCES
.........................................................................................................................
144
10
ABBREVATIONS
.....................................................................................................................
154
11
APPENDIX
............................................................................................................................
160
11.1
LIST
OF
TABLES
..............................................................................................................
160
11.2
PUBLICATIONS
................................................................................................................
162
12
DANKSAGUNG
......................................................................................................................
163
|
| any_adam_object | 1 |
| author | Lühr, Jennifer J. |
| author_GND | (DE-588)1181018145 |
| author_facet | Lühr, Jennifer J. |
| author_role | aut |
| author_sort | Lühr, Jennifer J. |
| author_variant | j j l jj jjl |
| building | Verbundindex |
| bvnumber | BV045515789 |
| ctrlnum | (OCoLC)1121486744 (DE-599)DNB1180440021 |
| discipline | Biologie |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV045515789 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T08:20:16Z |
| institution | BVB |
| language | German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-030900185 |
| oclc_num | 1121486744 |
| open_access_boolean | |
| owner | DE-29 DE-29T DE-355 DE-BY-UBR |
| owner_facet | DE-29 DE-29T DE-355 DE-BY-UBR |
| physical | IV, 164 Seiten Illustrationen, Diagramme 21 cm |
| publishDate | 2018 |
| publishDateSearch | 2018 |
| publishDateSort | 2018 |
| record_format | marc |
| spelling | Lühr, Jennifer J. Verfasser (DE-588)1181018145 aut Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane vorgelegt von Jennifer J. Lühr Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane Erlangen ; Nürnberg [2018] IV, 164 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2018 Lectine (DE-588)4034971-8 gnd rswk-swf Plasmamembran (DE-588)4067550-6 gnd rswk-swf Dendritische Zelle (DE-588)4313846-9 gnd rswk-swf Rezeptor (DE-588)4049722-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dendritische Zelle (DE-588)4313846-9 s Plasmamembran (DE-588)4067550-6 s Rezeptor (DE-588)4049722-7 s Lectine (DE-588)4034971-8 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030900185&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lühr, Jennifer J. Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane Lectine (DE-588)4034971-8 gnd Plasmamembran (DE-588)4067550-6 gnd Dendritische Zelle (DE-588)4313846-9 gnd Rezeptor (DE-588)4049722-7 gnd |
| subject_GND | (DE-588)4034971-8 (DE-588)4067550-6 (DE-588)4313846-9 (DE-588)4049722-7 (DE-588)4113937-9 |
| title | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| title_alt | Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| title_auth | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| title_exact_search | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| title_full | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane vorgelegt von Jennifer J. Lühr |
| title_fullStr | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane vorgelegt von Jennifer J. Lühr |
| title_full_unstemmed | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane vorgelegt von Jennifer J. Lühr |
| title_short | Dynamische Regulation und Kompartimentierung von C-Typ Lektinrezeptoren in der Zellmembran von dendritischen Zellen |
| title_sort | dynamische regulation und kompartimentierung von c typ lektinrezeptoren in der zellmembran von dendritischen zellen dynamic regulation and compartmentalization of c type lectin receptors in the dendritic cell membrane |
| title_sub | = Dynamic regulation and compartmentalization of C-type lectin receptors in the dendritic cell membrane |
| topic | Lectine (DE-588)4034971-8 gnd Plasmamembran (DE-588)4067550-6 gnd Dendritische Zelle (DE-588)4313846-9 gnd Rezeptor (DE-588)4049722-7 gnd |
| topic_facet | Lectine Plasmamembran Dendritische Zelle Rezeptor Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030900185&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT luhrjenniferj dynamischeregulationundkompartimentierungvonctyplektinrezeptoreninderzellmembranvondendritischenzellendynamicregulationandcompartmentalizationofctypelectinreceptorsinthedendriticcellmembrane AT luhrjenniferj dynamicregulationandcompartmentalizationofctypelectinreceptorsinthedendriticcellmembrane |