Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2007
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | V, 118 S. graph. Darst. 21 cm |
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MARC
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Record in the Search Index
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| adam_text | Table of contents
Abbreviations____________________________________________________________III
1 Introduction___________________________________________________________1
1.1 Tolerance____________________________________________________________1
1.1.1 Central tolerance_____________________________________________________________ 1
1.1.2 Peripheral T cell tolerance_____________________________________________________3
1.1.2.1 Passive (intrinsic) mechanisms of tolerance____________________________________3
1.1.2.2 Active (extrinsic) mechanisms of tolerance____________________________________5
1.1.3 Costimulation in the regulation of immune responses_________________________________7
1.2 Experimental autoimmune encephalomyelitis - a mouse model for multiple sclerosis 10
1.2.1 Myelin oligodendrocyte glycoprotein-specific TCR transgenic mice_____________________ 12
1.3 The role of DCs and B cells in tolerance induction ________________________________12
1.3.1 DCs in tolerance induction____________________________________________________ 12
1.3.2 B cells in tolerance induction__________________________________________________ 13
1.4 Objectives___________________________________________________________________16
2 Materials and Methods_________________________________________________18
2.1 Chemicals and biological material_________________________________________18
2.2 Molecular biology_____________________________________________________19
2.2.1 Preparation of genomic DNA__________________________________________________ 19
2.2.2 Polymerase chain reaction (PCR)_______________________________________________ 19
2.2.3 RNA isolation and Quantitative Real-Time PCR____________________________________21
2.2.4 Microarray analysis__________________________________________________________21
2.2.5 Isolation of plasmid DNA for southern probe preparation_____________________________22
2.2.6 Agarose gel electrophoresis and DNA gel extraction_________________________________22
2.2.7 Quantification of DNA and RNA_______________________________________________22
2.2.8 Southern blot analysis________________________________________________________22
23 Cell biology__________________________________________________________________23
2.3.1 Preparation of single cell suspensions from lymphoid organs__________________________23
2.3.2 Cell counting______________________________________________________________24
2.3.3 Flow cytometry_____________________________________________________________24
2.3.4 Magnetic cell sorting and FACS sorting__________________________________________26
2.3.5 CFSE labeling______________________________________________________________26
2.3.6 Culture of ex vivo lymphocytes_________________________________________________27
2.3.7 T and B cell activation_______________________________________________________27
2.3.8 Suppression assay___________________________________________________________28
2.3.9 Generation of bone marrow-derived DCs_________________________________________28
2.3.10 ELISA __________________________________________________________________28
2.4 Mouse experiments___________________________________________________________28
2.4.1 Mice________________________________________________________________________28
2.4.2 Induction of Cre recombinase activity and activation of APCs in vivo ___________________29
2.4.3 Adoptive transfer___________________________________________________________29
2.4.4 In vivo depletion of CD25* cells________________________________________________30
2.4.5 Induction and assessment of EAE_______________________________________________30
2.4.6 Statistics_________________________________________________________________30
3 Results_______________________________________________________________31
3.1 The IiMOG system____________________________________________________31
3.1.1 Functionality and specificity of the IiMOG system__________________________________33
3.2 The impact of MOG presentation by naive APCs on the course of EAE______________36
3.2.1 The role of DCs in induction of EAE____________________________________________37
3.2.2 The role of B cells in tolerance induction to EAE___________________________________39
33 The effect of MOG presentation by different APCs on MOG-spedfic CD4* T ceDs___42
3.3.1 Proliferation of MOG-specific T cells upon antigen encounter by different APCs___________42
332 Analysis of T cell proliferation upon presentation of MOG by resting and activated DCs_____47
I
Table of contents
3.3.3 Analysis of MOG-specific T cell activation by different APCs_________________________ 49
3.3.4 Expression of coinhibitory receptors after encounter of MOG-presenting B cells____________ 50
3.3.5 Functionality of B cell-triggered T cells___________________________________________51
3.3.6 In vivo restimulation leads to deletion of tolerized T cells_____________________________ 53
3.3.7 Analysis of expression of pro- and anti-apoptotic molecules upon interaction of T cells with
MOG-presenting B cells_______________________________________________________ 55
3.4 Analysis of BMOG/2D2 mice_____________________________________________________56
3.4.1 Presentation of MOG by B cells leads to deletion of MOG-specific T cells in the thymus_____56
3.4.2 Transgenic CD4* T cells in BMOG/2D2 mice are tolerized_____________________________ 58
3.5 Analysis of B cells in BMOG and BMOG/2D2 mice___________________________________60
3.5.1 Transfer of 2D2 T cells to BM0G mice does not affect the B cells _______________________ 60
3.5.2 Analysis of B cells in BMOG/2D2 mice____________________________________________ 61
3.6 The impact of B cells on regulatory T cells_______________________________________63
3.6.1 BMOG/2D2 mice exhibit an increased proportion of regulatory T cells_____________________ 63
3.6.2 Analysis of regulatory T cell marker expression by MOG-specific T cells upon encounter of
MOG-presenting B cells______________________________________________________65
3.6.3 Analysis of Foxp3 induction in MOG-specific T cells ^_________________________ 66
3.6.4 Induction of EAE upon depletion of regulatory T cells in BM xi mice 68
3.6.5 The role of B cells in the homeostasis of regulatory T cells____________________________ 69
3.7 Possible mechanisms of tolerance mediation by B cells____________________________71
3.7.1 Blockade of CD5 does not break tolerance of 2D2CD4T cells________________________71
3.7.2 PD-1 is not essential for B cell-mediated CD4+ T cell tolerance________________________71
3.7.3 Analysis of T cell proliferation upon blockade of CTLA-4____________________________ 74
3.8 Microarray analysis of tolerized MOG-specific CD4+ T cells________________________76
4 Discussion_____________________________________________________________77
4.1 The IiMOG system and its specificity____________________________________________78
4.2 Tolerance to EAE is induced by different APCs __________________________________79
A3 Characterization of tolerized MOG-specific T cells________________________________82
4.3.1 Tolerized MOG-specific T cells are deleted upon in vivo restimulation___________________ 84
4.3.2 Blockade of coinhibitory receptors to abrogate T cell tolerance_________________________ 85
4.4 B cells in central tolerance: Analysis of BMOG/2D2 mice____________________________87
4.5 Analysis of B cells in BMOG/2D2 mice_____________________________________________88
4.6 Does presentation of MOG by B cells affect the development of regulatory T cells?___88
4.7 The role of B cells in the homeostasis of Treg cells ________________________________91
5 Summary______________________________________________________________92
6 Zusammenfassung______________________________________________________93
7 References_____________________________________________________________95
8 Acknowledgements_____________________________________________________115
9 Versicherung__________________________________________________________116
10 Lebenslauf____________________________117
|
| any_adam_object | 1 |
| author | Frommer, Friederike 1976- |
| author_GND | (DE-588)134006127 |
| author_facet | Frommer, Friederike 1976- |
| author_role | aut |
| author_sort | Frommer, Friederike 1976- |
| author_variant | f f ff |
| building | Verbundindex |
| bvnumber | BV035212966 |
| ctrlnum | (OCoLC)222332529 (DE-599)BVBBV035212966 |
| dewey-full | 616.079EA |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 616 - Diseases |
| dewey-raw | 616.079ea |
| dewey-search | 616.079ea |
| dewey-sort | 3616.079 EA |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-09T21:28:44Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-017019169 |
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| physical | V, 118 S. graph. Darst. 21 cm |
| publishDate | 2007 |
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| spelling | Frommer, Friederike 1976- Verfasser (DE-588)134006127 aut Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen vorgelegt von Friedrike Frommer 2007 V, 118 S. graph. Darst. 21 cm txt rdacontent n rdamedia nc rdacarrier Köln, Univ., Diss., 2007 Toleranz Biologie (DE-588)4308830-2 gnd rswk-swf T-Lymphozyt (DE-588)4127387-4 gnd rswk-swf B-Zelle (DE-588)4147096-5 gnd rswk-swf Dendritische Zelle (DE-588)4313846-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content B-Zelle (DE-588)4147096-5 s Dendritische Zelle (DE-588)4313846-9 s Toleranz Biologie (DE-588)4308830-2 s T-Lymphozyt (DE-588)4127387-4 s DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017019169&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Frommer, Friederike 1976- Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen Toleranz Biologie (DE-588)4308830-2 gnd T-Lymphozyt (DE-588)4127387-4 gnd B-Zelle (DE-588)4147096-5 gnd Dendritische Zelle (DE-588)4313846-9 gnd |
| subject_GND | (DE-588)4308830-2 (DE-588)4127387-4 (DE-588)4147096-5 (DE-588)4313846-9 (DE-588)4113937-9 |
| title | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen |
| title_auth | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen |
| title_exact_search | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen |
| title_full | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen vorgelegt von Friedrike Frommer |
| title_fullStr | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen vorgelegt von Friedrike Frommer |
| title_full_unstemmed | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen vorgelegt von Friedrike Frommer |
| title_short | Exploring induction of T cell tolerance by B cells and dendritic cells upon conditional presentation of self antigen |
| title_sort | exploring induction of t cell tolerance by b cells and dendritic cells upon conditional presentation of self antigen |
| topic | Toleranz Biologie (DE-588)4308830-2 gnd T-Lymphozyt (DE-588)4127387-4 gnd B-Zelle (DE-588)4147096-5 gnd Dendritische Zelle (DE-588)4313846-9 gnd |
| topic_facet | Toleranz Biologie T-Lymphozyt B-Zelle Dendritische Zelle Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017019169&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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