The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response: = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Erlangen ; Nürnberg
2019
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | 138 Seiten Illustrationen, Diagramme |
Internformat
MARC
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| 041 | 0 | |a eng | |
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| 100 | 1 | |a Reimer, Dorothea |e Verfasser |0 (DE-588)1199410438 |4 aut | |
| 245 | 1 | 0 | |a The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response |b = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |c vorgelegt von Dorothea Reimer |
| 246 | 1 | 1 | |a Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| 264 | 1 | |a Erlangen ; Nürnberg |c 2019 | |
| 300 | |a 138 Seiten |b Illustrationen, Diagramme | ||
| 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 2019 | ||
| 650 | 0 | 7 | |a Immunreaktion |0 (DE-588)4133841-8 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Actin-Filament |0 (DE-588)4211337-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a B-Lymphozyt |0 (DE-588)4137121-5 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a B-Lymphozyt |0 (DE-588)4137121-5 |D s |
| 689 | 0 | 1 | |a Immunreaktion |0 (DE-588)4133841-8 |D s |
| 689 | 0 | 2 | |a Actin-Filament |0 (DE-588)4211337-4 |D s |
| 689 | 0 | |5 DE-604 | |
| 689 | 1 | 0 | |a Actin-Filament |0 (DE-588)4211337-4 |D s |
| 689 | 1 | 1 | |a B-Lymphozyt |0 (DE-588)4137121-5 |D s |
| 689 | 1 | 2 | |a Immunreaktion |0 (DE-588)4133841-8 |D s |
| 689 | 1 | |5 DE-604 | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1199117609/04 |3 Inhaltsverzeichnis |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-031629334 | ||
Datensatz im Suchindex
| _version_ | 1804180677901418496 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
1
SUMMARY
..........................................................................................................................
10
2
ZUSAMMENFASSUNG
.........................................................................................................
11
3
INTRODUCTION
.....................................................................................................................
13
3.1
THE
ADAPTIVE
IMMUNE
RESPONSE
....................................................................................
13
3.2
CENTRAL
STEPS
DURING
B
CELLS
DEVELOPMENT
AND
DIFFERENTIATION
.....................................
14
3.2.1
EARLY
B
CELL
MATURATION
IN
THE
BONE
MARROW
.............................................................
14
3.2.2
B
CELL
ACTIVATION
IN
SECONDARY
LYMPHOID
ORGANS
......................................................
15
3.2.3
THE
GERMINAL
CENTER
REACTION
...................................................................................
16
3.2.3.1
STRUCTURE
AND
CELLULAR
COMPONENTS
.......................................................................
16
3.
2.
3.2
IMMUNOGLOBULIN
ISOTYPE
SWITCHING
AND
AFFINITY
MATURATION
.................................
17
3.2.3.3
B
CELL
MIGRATION
WITHIN
GERMINAL
CENTERS
..............................................................
19
3.2.3.4
INTERACTION
BETWEEN
B
CELLS
AND
HELPER
CELLS
WITHIN
GERMINAL
CENTERS
................
20
3.2.4
TERMINAL
B
CELL
DIFFERENTIATION
INTO
PLASMA
CELLS
AND
MEMORY
B
CELLS
................... 23
3.2.5
DIFFERENTIAL
TRANSCRIPTIONAL
CONTROL
OF
GERMINAL
CENTER
B
CELLS
AND
PLASMA
CELLS
...
24
3.3
THE
CYTOSKELETON
IN
LYMPHOCYTES
...................................................................................26
3.3.1
STRUCTURAL
COMPOSITION
OF
THE
CORTICAL
ACTIN-CYTOSKELETON
......................................26
3.3.2
RECIPROCAL
MODULATION
OF
THE
CYTOSKELETON
AND
BCR
SIGNALING
............................
27
3.3.3
CYTOSKELETAL
FORCES
DURING
THE
GERMINAL
CENTER
RESPONSE
......................................29
3.3.3.1
MECHANICAL
FORCES
IN
CELL
MIGRATION
......................................................................
30
3.3.3.2
MECHANICAL
FORCES
IN
CELL-CELL
INTERACTION
AND
THE
FORMATION
OF
AN
IMMUNE
SYNAPSE
.................................................................................................................
31
3.4
THE
ADAPTOR
PROTEIN
EFHD2
.............................................................................................
33
3.4.1
EXPRESSION,
GENOMIC
LOCATION,
PROTEIN
STRUCTURE
....................................................
33
3.4.2
THE
ROLE
OF
EFHD2
IN
IMMUNE
CELLS
..........................................................................
35
3.4.3
ASSOCIATION
OF
EFHD2
WITH
THE
ACTIN
CYTOSKELETON
...................................................
36
4
OBJECTIVE
OF
THIS
WORK
....................................................................................................
38
5
RESULTS
.............................................................................................................................
39
5.1
EFHD2
EXPRESSION
IN
RESTING
AND
ACTIVATED
PRIMARY
B
CELLS
.........................................
39
5.2
CHARACTERIZATION
OF
EFHD2
KO
AND
WT
B
CELLS
IN
VITRO
FOR
THEIR
CYTOSKELETAL
PROPERTIES
........................................................................................................................
40
5.2.1
ANALYSIS
OF
B
CELL
SPREADING
.....................................................................................
40
5.2.2
ANALYSIS
OF
B
CELL
MOTILITY
...........................................................................................42
5.2.3
ANALYSIS
OF
INTEGRIN
EXPRESSION
AND
ICAM-1
BINDING
..............................................
45
5.3
CHARACTERIZATION
OF
THE
DIFFERENTIATION
OF
EFHD2
KO
AND
WT
B
CELLS
IN
VITRO
............47
VI
5.3.1
ANALYSIS
OF
GERMINAL
CENTER-LIKE
B
CELL
DEVELOPMENT
..............................................48
5.3.2
ANALYSIS
OF
PLASMA
BLAST
DEVELOPMENT
...................................................................
50
5.4
CHARACTERIZATION
OF
EFHD2
KO
AND
WT
B
CELL
MIGRATION
IN
VIVO
WITHIN
GERMINAL
CENTERS
............................................................................................................................
52
5.5
CHARACTERIZATION
OF
PROLIFERATION
OF
SPLENIC
B
CELL
SUBSETS
IN
EFHD2
KO
AND
WT
MICE54
5.6
CHARACTERIZATION
OF
THE
GERMINAL
RESPONSE
IN
EFHD2
KO
AND
WT
MICE
UPON
IMMUNIZATION
WITH
NP-KLH
...........................................................................................
56
5.6.1
VALIDATING
THE
DETECTION
OF
ANTIGEN-SPECIFIC
B
CELLS
BY
FLOW
CYTOMETRY
...................
56
5.6.2
ANALYSIS
OF
THE
GERMINAL
CENTER
B
CELL
RESPONSE
....................................................
57
5.6.3
ANALYSIS
OF
T
HELPER
CELLS
..........................................................................................60
5.6.4
ANALYSIS
OF
THE
PLASMA
BLAST/PLASMA
CELL
RESPONSE
................................................
61
5.6.5
ANALYSIS
OF
ANTIGEN-SPECIFIC
SERUM
IMMUNOGLOBULINS
.............................................65
5.6.6
ANALYSIS
OF
BLIMPL
EXPRESSION
IN
SPLENIC
B
CELL
POPULATIONS
.................................67
5.6.7
ANALYSIS
OF
NP-SPECIFIC
B
CELL
GENERATION
IN
THE
CONTEXT
OF
B
CELL
INTRINSIC
EFHD2
DEFICIENCY
..................................................................................................................
68
5.7
CHARACTERIZATION
OF
THE
INTERACTION
OF
EFHD2
KO
AND
WT
B
CELLS
IN
VIVO
WITH
HELPER
CELLS
................................................................................................................................
71
5.7.1
ANALYSIS
OF
SURFACE
EXPRESSION
OF
INTERACTION
MOLECULES
.......................................
72
5.7.2
ANALYSIS
OF
B-T
FH
CELL
INTERACTION
.............................................................................
73
5.7.3
ANALYSIS
OF
B
CELL-FDC
INTERACTION
...........................................................................
74
6
DISCUSSION
......................................................................................................................
76
6.1
EFHD2
LIMITS
DYNAMIC
REARRANGEMENT
OF
THE
CORTICAL
F-ACTIN
CYTOSKELETON
.................
76
6.2
EFHD2
MODULATES
CELL
INTRINSIC
PROPERTIES
THAT
DEFINE
AFFINITY-BASED
B
CELL
SELECTION
79
6.3 EFHD2
MODULATES
CELL
EXTRINSIC
PARAMETERS
THAT
DETERMINE
B
CELL
COMPETITIVENESS
.82
6.4
EFHD2
MODULATES
THE
FATE
OF
GC
B
CELLS
.....................................................................
84
6.5
CONCLUDING
REMARKS
......................................................................................................
87
7
MATERIAL
............................................................................................................................
88
7.1
CONSUMABLES
.................................................................................................................
88
7.2
COMMERCIAL
KITS
.............................................................................................................
88
7.3
CHEMICALS
AND
REAGENTS
...............................................................................................
89
7.4
BUFFERS,
SOLUTIONS
AND
MEDIA
.........................................................................................
90
7.5
OLIGONUCLEOTIDES
............................................................................................................
91
7.6
ANTIBODIES
AND
FLUORESCENT
DYES
..................................................................................
91
7.7
INSTRUMENTS
AND
DEVICES
...............................................................................................
93
7.8
SOFTWARE
.........................................................................................................................
94
7.9
MOUSE
STRAINS
................................................................................................................
95
VII
8
METHODS
...........................................................................................................................
96
8.1
ANIMAL
HOUSING
...............................................................................................................
96
8.2
GENOTYPING
OF
MICE
........................................................................................................
96
8.2.1
DNA
EXTRACTION
FROM
EAR
AND
TAIL
BIOPSIES
...............................................................96
8.2.2
PCR
REACTIONS
...........................................................................................................
96
8.2.3
GEL
ELECTROPHORESIS
OF
PCR
PRODUCTS
......................................................................
98
8.3
INTRAPERITONEAL
IMMUNIZATION
OF
MICE
............................................................................
98
8.3.1
(4-HYDROXY-3-NITROPHENYL)ACETYL-KEYHOLE
LIMPET
HEMOCYANIN
(NP-KLH)
............
98
8.3.2
SHEEP
RED
BLOOD
CELLS
(SRBCS)
...............................................................................98
8.3.3
COLLECTION
OF
BLOOD
SAMPLES
FOR
SERUM
....................................................................
99
8.4
IN
VIVO
EDLL
APPLICATION
FOR
PROLIFERATION
ANALYSIS
.........................................................
99
8.5
PREPARATION
OF
PRIMARY
MURINE
CELLS
EX
VIVO
.................................................................
99
8.6
DETERMINATION
OF
CELL
NUMBERS
.....................................................................................
100
8.7
PURIFICATION
OF
NAIVE
B
CELLS
FROM
THE
SPLEEN
..............................................................
101
8.8
ADOPTIVE
B
CELL
TRANSFER
INTO
LY5.1
MICE
.....................................................................
101
8.9
CULTIVATION
OF
PRIMARY
MURINE
B
CELLS
IN
VITRO
.............................................................
102
8.9.1
B
CELL
ACTIVATION
AND
SHORT
TERM
CULTIVATION
............................................................
102
8.9.2
B
CELL
CULTIVATION
USING
THE
IGB
SYSTEM
..................................................................
102
8.10
FLOW
CYTOMETRIC
ANALYSIS
..............................................................................................
103
8.10.1
STAINING
OF
SURFACE
PROTEINS
...................................................................................
103
8.10.2
COMMON
GATING
STRATEGY
FOR
VIABLE
CELLS
AND
EXCLUSION
OF
AUTOFLUORESCENCE
....
104
8.10.3
FLOW
CYTOMETRIC
DETECTION
OF
EDLL
POSITIVE
CELLS
....................................................
105
8.11
ANALYSIS
OF
SERUM
IMMUNOGLOBULINS
WITH
ELISA
.......................................................
105
8.12
IN
VITRO
MIGRATION
ASSAY
OF
ACTIVATED
B
CELLS
...............................................................
106
8.12.1
SAMPLE
PREPARATION
AND
ASSAY
SETUP
....................................................................
106
8.12.2
SPINNING
DISC
CONFOCAL
MICROSCOPY
AND
QUANTIFICATION
.........................................
107
8.13
IN
VITRO
SPREADING
ASSAY
OF
ACTIVATED
B
CELLS
..............................................................
107
8.13.1
SAMPLE
PREPARATION
AND
ASSAY
SETUP
....................................................................
107
8.13.2
LASER
SCANNING
MICROSCOPY
AND
QUANTIFICATION
.....................................................
108
8.14
IN
VIVO
B
CELL
MIGRATION
ANALYSIS
..................................................................................
108
8.14.1
CELL
PREPARATION
AND
ASSAY
SET
UP
..........................................................................
108
8.14.2
TWO-PHOTON
MICROSCOPY
..........................................................................................
109
8.14.3
QUANTIFICATION
OF
B
CELL
MIGRATION
AND
B-FDC
INTERACTION
.....................................
109
8.15
STATISTICAL
ANALYSIS
........................................................................................................
110
9
APPENDIX
........................................................................................................................
111
9.1
ABBREVIATION
INDEX
....................................................................................................
111
VIII
9.2
INDEX
OF
UNITS,
PREFIXES
AND
GREEK
LETTERS
..................................................................
114
9.3
FIGURE
INDEX
..................................................................................................................
115
9.4
TABLE
INDEX
...................................................................................................................
117
9.5
REFERENCE
INDEX
..........................................................................................................
118
9.6
PUBLICATION
INDEX
..........................................................................................................
138
IX
|
| any_adam_object | 1 |
| author | Reimer, Dorothea |
| author_GND | (DE-588)1199410438 |
| author_facet | Reimer, Dorothea |
| author_role | aut |
| author_sort | Reimer, Dorothea |
| author_variant | d r dr |
| building | Verbundindex |
| bvnumber | BV046251116 |
| ctrlnum | (OCoLC)1128845356 (DE-599)DNB1199117609 |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-10T08:39:34Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-031629334 |
| oclc_num | 1128845356 |
| 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 | 138 Seiten Illustrationen, Diagramme |
| publishDate | 2019 |
| publishDateSearch | 2019 |
| publishDateSort | 2019 |
| record_format | marc |
| spelling | Reimer, Dorothea Verfasser (DE-588)1199410438 aut The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort vorgelegt von Dorothea Reimer Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort Erlangen ; Nürnberg 2019 138 Seiten Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2019 Immunreaktion (DE-588)4133841-8 gnd rswk-swf Actin-Filament (DE-588)4211337-4 gnd rswk-swf B-Lymphozyt (DE-588)4137121-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content B-Lymphozyt (DE-588)4137121-5 s Immunreaktion (DE-588)4133841-8 s Actin-Filament (DE-588)4211337-4 s DE-604 B:DE-101 application/pdf https://d-nb.info/1199117609/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031629334&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Reimer, Dorothea The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort Immunreaktion (DE-588)4133841-8 gnd Actin-Filament (DE-588)4211337-4 gnd B-Lymphozyt (DE-588)4137121-5 gnd |
| subject_GND | (DE-588)4133841-8 (DE-588)4211337-4 (DE-588)4137121-5 (DE-588)4113937-9 |
| title | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| title_alt | Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| title_auth | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| title_exact_search | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| title_full | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort vorgelegt von Dorothea Reimer |
| title_fullStr | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort vorgelegt von Dorothea Reimer |
| title_full_unstemmed | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort vorgelegt von Dorothea Reimer |
| title_short | The role of EFhd2/Swiprosin-1 for actin dynamics in B lymphocytes and its function in the germinal center dependent immune response |
| title_sort | the role of efhd2 swiprosin 1 for actin dynamics in b lymphocytes and its function in the germinal center dependent immune response die rolle von efhd2 swiprosin 1 fur die dynamik des aktin zytoskeletts in b lymphozyten und seine funktion in der keimzentrums abhangigen immunantwort |
| title_sub | = Die Rolle von EFhd2/Swiprosin-1 für die Dynamik des Aktin-Zytoskeletts in B-Lymphozyten und seine Funktion in der Keimzentrums-abhängigen Immunantwort |
| topic | Immunreaktion (DE-588)4133841-8 gnd Actin-Filament (DE-588)4211337-4 gnd B-Lymphozyt (DE-588)4137121-5 gnd |
| topic_facet | Immunreaktion Actin-Filament B-Lymphozyt Hochschulschrift |
| url | https://d-nb.info/1199117609/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031629334&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT reimerdorothea theroleofefhd2swiprosin1foractindynamicsinblymphocytesanditsfunctioninthegerminalcenterdependentimmuneresponsedierollevonefhd2swiprosin1furdiedynamikdesaktinzytoskelettsinblymphozytenundseinefunktioninderkeimzentrumsabhangigenimmunantwort AT reimerdorothea dierollevonefhd2swiprosin1furdiedynamikdesaktinzytoskelettsinblymphozytenundseinefunktioninderkeimzentrumsabhangigenimmunantwort |
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