Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2008
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| Subjects: | |
| Online Access: | Volltext http://d-nb.info/989029778/34 Volltext Inhaltsverzeichnis |
| Physical Description: | 104 S. Ill., graph. Darst |
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MARC
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| adam_text | Table of contents
A. Summary 1
A. Zusammenfassung 2
B. Indroduction 3
B.l. The Immune System 3
B. 1.1. Innate immunity 3
B. 1.2. Adaptive immunity 4
B.I.2.1.Bcells 5
B.l.2.2. T-cells 6
B.2. Dendritic cells 8
B.2.1. Dendritic cell subsets 9
B.2.2. Dendritic Cell Life Cycle 10
B.2.3. DC and anti-tumor immunotherapy 12
B.3. Oncolytic adenoviruses 15
B.3.1 Adenoviruses 15
B.3.2. Oncolytic adenoviruses 16
B.4. Investigation of anti-tumor immune responses induced by adenoviral oncolysis 19
B.S. Objectives 20
C. Materials and methods 21
C.I. Materials 21
C. 1 1 Eukaryotic cell lines 21
C. 1.2. Medium and solutions for cell culture 21
C.1.3. Nucleic acids 22
C.l.3.1. Oligonucleotids used for Real-Time PCR 22
C.l.3.2. Oligonucleotids used for PCR 22
C.I.4. Antibodies 22
C.I.5. Chemicals and enzymes 23
C. 1.6. Buffers and solution 24
C. 1.6.1. General buffers and solutions 24
C. 1.6.2. Electrophoresis of nucleid acids 24
C. 1.6.3. Electrophoresis of proteins 24
C.2. Methods 25
C.2.1. Methods for detection of protein 25
C.2.1.1. Generating cell lysate 25
C.2.1.2. Determination of protein concentration with Bio-Rad 25
C.2.1.3. Denaturing Discontinuous Polyacrylamide Gel Electrophoresis according to Laemmli 26
C.2.1.4. Western blotting of proteins 26
C.2.1.5. Detection of poly (ADP-ribose) polymerase (PARP) 26
C.2.2. Cell culture and generation of DCs and T cells 27
C.2.3. Recombinant adenoviruses ____ 28
C.2.4. Luciferase assay 29
C.2.5. FACS analysis of GFP expression, cell surface staining and Tetramer staining. 30
C.2.6. Cytotoxicity assays 30
C.2.7. Quantification of virus genomes by real-time PCR 31
C.2.8. Cytotoxic T cell induction assay. 32
C.2.9. IL-12 production by DCs. ~ 32
C.2.10. Cytometric Bead Array (CBA) Z 33
C.2.11. Analysis of cell death of melanoma cells ~ ~ ~ 33
C.2.12. Labeling of melanoma cells 34
C.2.13. Phagocytosis assay __ 35
C.2.14. Migration assay_ ~ ~ ~___^_ 35
D. Results 36
D.I. Influence of oncolytic adenoviruses on DC biology 36
D. 1.1. Recombinant adenoviruses _____ _____ ______ ______________________ 36
D. 1.2. Adenoviral transduction of immature and mature DCs ________ 37
D. 1.3. Replication and cytotoxicity of oncolytic adenoviruses in iDC and mDC cultures. 39
D.I.4. Expression of MHC molecules, costimulatory molecules and secretion of cytokines by DCs after
infection with oncolytic adenoviruses. _________„ 43
D. 1.5. Modulation of MC-induced expression of MHCs, costimulatory molecules and cytokines by DCs
after oncolytic adenovirus infection 47
D.I.6. T cell stimulation by DCs after infection with oncolytic adenoviruses 48
D.I.7. Modulation of 1L-12 production by DCs after oncolytic adenovirus infection. 50
D.2. Analysis of oncolytic adenovirus infected melanoma cells and their influence on DC
biology 52
D.2.1. Characterization of oncolytic adenovirus induced melanoma cell death 52
D.2.2. Analysis of cytokine production by oncolysate 57
D.2.3. Establishment of a phagocytosis assay for DCs and oncolysate 58
D.2.4. Uptake of oncolysate by DCs __________ 59
D.2.5. Phenotype of DCs which phagocytosed oncolysate _^_______________________.______ 61
D.2.6. Migration capacity of DCs after phagocytosis of oncolysate 62
D.2.7. Phenotype of DCs after uptake of oncolysate expressing CD40L 64
F. Discussion 68
F.I Modulation of DC biology by oncolytic adenoviruses 68
F.2 Mode of cell death in melanoma cells infected by oncolytic adenoviruses 72
F.3 Modulation of DC biology by melanoma cells lysed by oncolytic adenoviruses 74
F.4. Outlook 79
F5. Conclusions 83
G. Abbreviations 85
H. References 88
I. Personal publications 102
1.1. Original scientific publications 102
1.2. Poster presentations 102
1.3. Participation in the trainee program of graduate college 592 102
J. Curriculum Vitae 103
K. Acknowledgements/Danksagung 104
|
| adam_txt |
Table of contents
A. Summary 1
A. Zusammenfassung 2
B. Indroduction 3
B.l. The Immune System 3
B. 1.1. Innate immunity 3
B. 1.2. Adaptive immunity 4
B.I.2.1.Bcells 5
B.l.2.2. T-cells 6
B.2. Dendritic cells 8
B.2.1. Dendritic cell subsets 9
B.2.2. Dendritic Cell Life Cycle 10
B.2.3. DC and anti-tumor immunotherapy 12
B.3. Oncolytic adenoviruses 15
B.3.1 Adenoviruses 15
B.3.2. Oncolytic adenoviruses 16
B.4. Investigation of anti-tumor immune responses induced by adenoviral oncolysis 19
B.S. Objectives 20
C. Materials and methods 21
C.I. Materials 21
C. 1 1 Eukaryotic cell lines 21
C. 1.2. Medium and solutions for cell culture 21
C.1.3. Nucleic acids 22
C.l.3.1. Oligonucleotids used for Real-Time PCR 22
C.l.3.2. Oligonucleotids used for PCR 22
C.I.4. Antibodies 22
C.I.5. Chemicals and enzymes 23
C. 1.6. Buffers and solution 24
C. 1.6.1. General buffers and solutions 24
C. 1.6.2. Electrophoresis of nucleid acids 24
C. 1.6.3. Electrophoresis of proteins 24
C.2. Methods 25
C.2.1. Methods for detection of protein 25
C.2.1.1. Generating cell lysate 25
C.2.1.2. Determination of protein concentration with Bio-Rad 25
C.2.1.3. Denaturing Discontinuous Polyacrylamide Gel Electrophoresis according to Laemmli 26
C.2.1.4. Western blotting of proteins 26
C.2.1.5. Detection of poly (ADP-ribose) polymerase (PARP) 26
C.2.2. Cell culture and generation of DCs and T cells 27
C.2.3. Recombinant adenoviruses _ 28
C.2.4. Luciferase assay 29
C.2.5. FACS analysis of GFP expression, cell surface staining and Tetramer staining. 30
C.2.6. Cytotoxicity assays 30
C.2.7. Quantification of virus genomes by real-time PCR 31
C.2.8. Cytotoxic T cell induction assay. 32
C.2.9. IL-12 production by DCs. ~ 32
C.2.10. Cytometric Bead Array (CBA) ' ' " Z 33
C.2.11. Analysis of cell death of melanoma cells ~ ~ ~ 33
C.2.12. Labeling of melanoma cells 34
C.2.13. Phagocytosis assay _ 35
C.2.14. Migration assay_ ~ ~ ' ~_^_ 35
D. Results 36
D.I. Influence of oncolytic adenoviruses on DC biology 36
D. 1.1. Recombinant adenoviruses _ _ _ _ 36
D. 1.2. Adenoviral transduction of immature and mature DCs _ 37
D. 1.3. Replication and cytotoxicity of oncolytic adenoviruses in iDC and mDC cultures. 39
D.I.4. Expression of MHC molecules, costimulatory molecules and secretion of cytokines by DCs after
infection with oncolytic adenoviruses. _„ 43
D. 1.5. Modulation of MC-induced expression of MHCs, costimulatory molecules and cytokines by DCs
after oncolytic adenovirus infection 47
D.I.6. T cell stimulation by DCs after infection with oncolytic adenoviruses 48
D.I.7. Modulation of 1L-12 production by DCs after oncolytic adenovirus infection. 50
D.2. Analysis of oncolytic adenovirus infected melanoma cells and their influence on DC
biology 52
D.2.1. Characterization of oncolytic adenovirus induced melanoma cell death 52
D.2.2. Analysis of cytokine production by oncolysate 57
D.2.3. Establishment of a phagocytosis assay for DCs and oncolysate 58
D.2.4. Uptake of oncolysate by DCs _ 59
D.2.5. Phenotype of DCs which phagocytosed oncolysate _^_._ 61
D.2.6. Migration capacity of DCs after phagocytosis of oncolysate 62
D.2.7. Phenotype of DCs after uptake of oncolysate expressing CD40L 64
F. Discussion 68
F.I Modulation of DC biology by oncolytic adenoviruses 68
F.2 Mode of cell death in melanoma cells infected by oncolytic adenoviruses 72
F.3 Modulation of DC biology by melanoma cells lysed by oncolytic adenoviruses 74
F.4. Outlook 79
F5. Conclusions 83
G. Abbreviations 85
H. References 88
I. Personal publications 102
1.1. Original scientific publications 102
1.2. Poster presentations 102
1.3. Participation in the trainee program of graduate college 592 102
J. Curriculum Vitae 103
K. Acknowledgements/Danksagung 104 |
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| spelling | Schierer, Stephan Reinhard Verfasser aut Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses von Stephan Reinhard Schierer 2008 104 S. Ill., graph. Darst txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2008 Langzeitarchivierung Nationalbibliothek gewährleistet Onkologie (DE-588)4075658-0 gnd rswk-swf Adenoviren (DE-588)4141412-3 gnd rswk-swf Therapie (DE-588)4059798-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Onkologie (DE-588)4075658-0 s Therapie (DE-588)4059798-2 s Adenoviren (DE-588)4141412-3 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:29-opus-9207 https://open.fau.de/handle/openfau/624 Verlag kostenfrei Volltext http://d-nb.info/989029778/34 https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-9207 Verlag kostenfrei Volltext HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016498110&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schierer, Stephan Reinhard Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses Onkologie (DE-588)4075658-0 gnd Adenoviren (DE-588)4141412-3 gnd Therapie (DE-588)4059798-2 gnd |
| subject_GND | (DE-588)4075658-0 (DE-588)4141412-3 (DE-588)4059798-2 (DE-588)4113937-9 |
| title | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| title_auth | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| title_exact_search | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| title_exact_search_txtP | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| title_full | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses von Stephan Reinhard Schierer |
| title_fullStr | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses von Stephan Reinhard Schierer |
| title_full_unstemmed | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses von Stephan Reinhard Schierer |
| title_short | Modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| title_sort | modulation of dendritic cell biology by oncolytic adenoviruses and by melanoma cells lysed by oncolytic adenoviruses |
| topic | Onkologie (DE-588)4075658-0 gnd Adenoviren (DE-588)4141412-3 gnd Therapie (DE-588)4059798-2 gnd |
| topic_facet | Onkologie Adenoviren Therapie Hochschulschrift |
| url | https://open.fau.de/handle/openfau/624 http://d-nb.info/989029778/34 https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-9207 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016498110&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schiererstephanreinhard modulationofdendriticcellbiologybyoncolyticadenovirusesandbymelanomacellslysedbyoncolyticadenoviruses |