Genetic modification of dendritic cells for antigen specific stimulation of t cells:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2004
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | VII, 87 Bl. graph. Darst. 30 cm |
Staff View
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| 245 | 1 | 0 | |a Genetic modification of dendritic cells for antigen specific stimulation of t cells |c von Tatjana Clarissa Gust |
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| 502 | |a Berlin, Humboldt-Univ., Diss.,2004 | ||
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Record in the Search Index
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| adam_txt |
TABLE
OF
CONTENTS
TABLE
OF
CONTENTS
ZUSAMMENFASSUNG
.
I
ABSTRACT
.
II
TABLE
OF
CONTENTS
.
ILL
ABBREVIATIONS
.
VI
1.
INTRODUCTION
.
1
1.1.
IMMUNOTHERAPY
.
1
1.2.
DC
AND
THEIR
ROLE
IN
THE
IMMUNE
SYSTEM
.
2
1.3.
GENERATION
AND
CHARACTERIZATION
OF
DC
SUBTYPES
AND
POPULATIONS
.
5
1.4.
DC
IN
IMMUNOTHERAPY
.
6
1.5.
GENE
TRANSFER
INTO
DC
.
8
1.5.1.
RECEPTOR-MEDIATED
GENE
DELIVERY
INTO
DC
WITH
ADENOVIRUS
AS
LIGAND
.
8
1.5.2.
LENTIVIRAL
VECTORS
FOR
GENE
DELIVERY
INTO
DC
.
10
1.6.
OBJECTIVES
.
12
2.
MATERIALS
AND
METHODS
.
13
2.1.
MATERIALS
.
13
2.1.1.
APPLIANCES
AND
COMPUTER
SOFTWARE
.
13
2.1.2.
CHEMICALS,
SOLUTIONS
AND
BUFFERS
.
13
2.1.3.
ENZYMES
.
14
2.1.4.
ANTIBODIES
.
14
2.1.4.1.
CHARACTERIZATION
OF
HUMAN
CELLS
.
14
2.1.4.2.
CHARACTERIZATION
OF
MOUSE
CELLS
.
15
2.1.4.3.
IMMUNOMAGNETIC
BEAD
PURIFICATION
.
15
2.1.5.
OLIGONUCLEOTIDES
.
15
2.1.6.
PEPTIDES
.
15
2.1.7.
BACTERIAL
STRAINS
.
16
2.1.8.
PLASMIDS
.
16
2.1.8.1.
PLASMIDS
FOR
REPORTER
GENE
ASSAYS
.
16
2.1.8.2.
OVALBUMIN
PLASMIDS
.
16
2.1.8.3.
LENTIVIRAL
PLASMIDS
.
17
2.1.9.
MOUSE
STRAINS
.
18
2.1.10.
CELL
LINES
.
18
2.1.11.
CELL
CULTURE
MEDIA
AND
ADDITIVES
.
18
2.1.12.
CYTOKINES
AND
GROWTH
FACTORS
.
18
2.1.12.1.
CYTOKINES
FOR
HUMAN
MONOCYTE-DERIVED
DC
.
18
2.1.12.2.
CYTOKINES
FOR
MOUSE
BONE
MARROW-DERIVED
DC
.
19
TABLE
OF
CONTENTS
2.2.
METHODS
.
19
2.2.1.
CONSTRUCTION
OF
PGEM4Z/TFR-OVA/A64
AND
PGEM4Z/II-OVAI+II/A64
.
19
2.2.2.
GENERATION
OF
RECOMBINANT
ADENOVIRAL
PLASMIDS
.
19
2.2.3.
CONSTRUCTION
OF
LENTIVIRAL
TRANSFER
PLASMIDS
.
20
2.2.4.
IN
VITRO
TRANSCRIPTION
.
20
2.2.5.
PRODUCTION
OF
RECOMBINANT
ADENOVIRAL
AND
LENTIVIRAL
VECTORS
.
21
2.2.5.1.
GENERATION
OF
HIGH
TITER
ADENOVIRUS
STOCKS
.
21
2.2.5.2.
GENERATION
OF
LENTIVIRAL
VECTORS
.
21
2.2.6.
DC
PREPARATION
AND
CULTURE
.
22
2.2.6.1.
ISOLATION
AND
CULTURE
OF
HUMAN
MONOCYTE-DERIVED
DC
.
22
2.2.6.2.
ISOLATION
AND
CULTURE
OF
MOUSE
BONE
MARROW-DERIVED
DC
.
23
2.2.7.
CHARACTERIZATION
OF
DC
BY
FLOW
CYTOMETRY
.
23
2.2.7.1.
HUMAN
MONOCYTE-DERIVED
DC
.
23
2.2.7.2.
MOUSE
BONE
MARROW-DERIVED
DC
.
23
2.2.8.
TRANSFECTION
PROTOCOLS
.
24
2.2.8.1.
TRANSFECTION
OF
AD/PEI/DNA
OR
AD/PEI/RNA
COMPLEXES
.
24
2.2.8.2.
TRANSFECTION
OF
AD/POLYBRENE/DNA
OR
AD/POLYBRENE/RNA
COMPLEXES
.
24
2.2.8.3.
NUCLEOFECTION
.
25
2.2.8.4.
ADENOVIRAL
INFECTIONS
.
25
2.2.8.5.
LENTIVIRAL
INFECTIONS
.
25
2.2.9.
EVALUATION
OF
TRANSFECTION
EFFICIENCY
.
25
2.2.10.
ANTIGEN-SPECIFIC
T
CELL
STIMULATION
.
26
2.2.10.1.
ISOLATION
OF
PRIMARY
MURINE
TRANSGENIC
T
CELLS
.
26
2.2.10.2.
T
CELL
PROLIFERATION
ASSAY
.
26
2.2.10.3.
DETECTION
OF
ANTIGEN-SPECIFIC
IL-2
PRODUCTION
.
26
3.
RESULTS
.
27
3.1.
RNA
VERSUS
DNA
TRANSDUCTION
OF
DC
FOR
TRANSIENT
TRANSGENE
EXPRESSION
.
27
3.1.1.
TRANSFECTION
OF
K562
CELLS
WITH
AD/PEI/DNA
OR
AD/PEI/RNA
COMPLEXES
.
27
3.1.2.
TRANSFECTION
OF
MOUSE
DC
WITH
AD/PEI/DNA
OR
AD/PEI/RNA
COMPLEXES
.
30
3.1.3.
AD/PEI
TRANSDUCTION
OF
MURINE
DC
AND
OVA-SPECIFIC
T
CELL
STIMULATION
.
33
3.1.3.1.
RNA
TRANSDUCED
DC
ARE
POTENT
STIMULATORS
OF
OVA-SPECIFIC
CD8
+
T
CELLS
.
33
3.1.3.2.
INACTIVATED
VERSUS
ACTIVATED
ADENOVIRUS
PARTICLES
IN
AD/PEI/RNA
TRANSFECTION
.
35
3.1.3.3.
IMPACT
OF
CAPPING
AND
POLYADENYLATION
OF
RNA
ON
TRANSGENE
EXPRESSION
AND
T
CELL
STIMULATION
.
36
3.1.4.
EFFECT
OF
COXSACKIE-ADENOVIRUS
RECEPTOR
EXPRESSION
ON
AD/PEI
TRANSFECTION
OF
DC
AND
T
CELL
STIMULATION
.
40
3.1.5.
AD/PEI
TRANSFECTION
OF
HUMAN
MONOCYTE-DERIVED
DC
.
43
3.1.6.
ALTERNATIVE
TRANSFECTION
SYSTEMS
FOR
TRANSIENT
TRANSFECTION
OF
DC
.
44
3.1.6.1.
AD/POLYBRENE
TRANSFECTION
OF
HUMAN
AND
MOUSE
DC
.
44
3.1.6.2.
NUCLEOFECTION
OF
HUMAN
MONOCYTE-DERIVED
DC
.
46
3.2.
LENTIVIRAL
TRANSDUCTION
OF
DC
FOR
STABLE
TRANSGENE
EXPRESSION
.
48
3.2.1.
LENTIVIRAL
TRANSDUCTION
OF
MOUSE
BONE
MARROW-DERIVED
DC
.
48
TABLE
OF
CONTENTS
3.2.2.
ACTIVITY
OF
CENTRIFUGATION,
POLYBRENE
AND
PROTAMINE
SULFATE
ON
LENTIVIRAL
TRANSDUCTION
OF
DC
.
50
3.2.3.
DURATION
OF
TRANSGENE
EXPRESSION
IN
DC
TRANSDUCED
WITH
LENTIVIRAL
VECTORS
.
52
3.2.4.
LENTIVIRAL
TRANSDUCED
DC
INDUCE
CD8
+
AND
CD4
+
T
CELL
RESPONSES
.
53
4.
DISCUSSION
.
55
4.1.
TRANSIENT
TRANSDUCTION
OF
DC
.
55
4.1.1.
AD/PEI/RNA
VERSUS
AD/PEI/DNA
TRANSDUCTION
OF
DC
.
55
4.1.2.
COXSACKIE-ADENOVIRUS
RECEPTOR
EXPRESSION
AND
AD/PEI
TRANSDUCTION
OF
DC
.
58
4.1.3.
AD/POLYBRENE
TRANSFECTION
OF
DC
.
59
4.2.
STABLE
TRANSDUCTION
OF
DC
BY
LENTIVIRAL
VECTORS
.
60
REFERENCES
.
62
APPENDIX
.
79
PLASMIDS
.
79
CURRICULUM
VITAE
.
83
PUBLICATIONS
.
84
POSTERS
.
85
ORAL
PRESENTATIONS
.
85
DANKSAGUNGEN
.
86
EIDESSTATTLICHE
ERKLARUNG
.
87 |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV023123916 |
| illustrated | Illustrated |
| index_date | 2024-07-02T19:52:37Z |
| indexdate | 2024-07-09T21:11:36Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-016326351 |
| oclc_num | 634778353 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-11 |
| owner_facet | DE-355 DE-BY-UBR DE-11 |
| physical | VII, 87 Bl. graph. Darst. 30 cm |
| publishDate | 2004 |
| publishDateSearch | 2004 |
| publishDateSort | 2004 |
| record_format | marc |
| spelling | Gust, Tatjana Clarissa 1975- Verfasser (DE-588)129634255 aut Genetic modification of dendritic cells for antigen specific stimulation of t cells von Tatjana Clarissa Gust 2004 VII, 87 Bl. graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Berlin, Humboldt-Univ., Diss.,2004 T-Lymphozyt (DE-588)4127387-4 gnd rswk-swf Dendritische Zelle (DE-588)4313846-9 gnd rswk-swf Transduktion (DE-588)4333506-8 gnd rswk-swf Aktivierung Physiologie (DE-588)4243777-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content T-Lymphozyt (DE-588)4127387-4 s Aktivierung Physiologie (DE-588)4243777-5 s Dendritische Zelle (DE-588)4313846-9 s Transduktion (DE-588)4333506-8 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016326351&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gust, Tatjana Clarissa 1975- Genetic modification of dendritic cells for antigen specific stimulation of t cells T-Lymphozyt (DE-588)4127387-4 gnd Dendritische Zelle (DE-588)4313846-9 gnd Transduktion (DE-588)4333506-8 gnd Aktivierung Physiologie (DE-588)4243777-5 gnd |
| subject_GND | (DE-588)4127387-4 (DE-588)4313846-9 (DE-588)4333506-8 (DE-588)4243777-5 (DE-588)4113937-9 |
| title | Genetic modification of dendritic cells for antigen specific stimulation of t cells |
| title_auth | Genetic modification of dendritic cells for antigen specific stimulation of t cells |
| title_exact_search | Genetic modification of dendritic cells for antigen specific stimulation of t cells |
| title_exact_search_txtP | Genetic modification of dendritic cells for antigen specific stimulation of t cells |
| title_full | Genetic modification of dendritic cells for antigen specific stimulation of t cells von Tatjana Clarissa Gust |
| title_fullStr | Genetic modification of dendritic cells for antigen specific stimulation of t cells von Tatjana Clarissa Gust |
| title_full_unstemmed | Genetic modification of dendritic cells for antigen specific stimulation of t cells von Tatjana Clarissa Gust |
| title_short | Genetic modification of dendritic cells for antigen specific stimulation of t cells |
| title_sort | genetic modification of dendritic cells for antigen specific stimulation of t cells |
| topic | T-Lymphozyt (DE-588)4127387-4 gnd Dendritische Zelle (DE-588)4313846-9 gnd Transduktion (DE-588)4333506-8 gnd Aktivierung Physiologie (DE-588)4243777-5 gnd |
| topic_facet | T-Lymphozyt Dendritische Zelle Transduktion Aktivierung Physiologie Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016326351&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gusttatjanaclarissa geneticmodificationofdendriticcellsforantigenspecificstimulationoftcells |