IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
Mensch & Buch Verl.
2011
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 81 S. Ill., graph. Darst. |
| ISBN: | 9783866649545 |
Internformat
MARC
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| 245 | 1 | 0 | |a IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis |c Markus Zoller |
| 264 | 1 | |a Berlin |b Mensch & Buch Verl. |c 2011 | |
| 300 | |a 81 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |a Zugl.: Berlin, Humboldt-Univ., Diss., 2011 | ||
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| 650 | 0 | 7 | |a Immunglobulin G |0 (DE-588)4127637-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Autoantikörper |0 (DE-588)4215429-7 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Plasmazelle |0 (DE-588)4174842-6 |2 gnd |9 rswk-swf |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-022562248 | ||
Datensatz im Suchindex
| _version_ | 1804145706524475392 |
|---|---|
| adam_text | IMAGE 1
IV TABLE OF CONTENTS
I ZUSAMMENFASSUNG I
II SUMMARY II
III ABBREVIATIONS ILL
IV TABLE OF CONTENTS VI
1 INTRODUCTION 1
1.1 THE IMMUNE SYSTEM 1
1.2 IMMUNOGLOBULINS 1
1.3 GENERATION OF DIVERSITY IN THE IMMUNO GLOBULIN REPERTOIRE 3
1.4 GENERATION OF MEMORY B CELLS AND PLASMA CELLS 4
1.5 TOLERANCE AND SELF-REACTIVITY IN HUMANS 6
1.6 AUTOIMMUNITY 7
1.7 SYSTEMIC SCLEROSIS (SSC) 8
1.8 B CELLS AND AUTOANTIBODIES IN SSC 9
2 AIM 11
3 METHODS 12
3.1 STRATEGY OUTLINE 12
3.2 FLUORESCENCE-ACTIVATED CELL SORTING (FACS) 13
3.3 REVERSE TRANSCRIPTION - POLYMERASE CHAIN REACTION (RT-PCR) 14
3.3.1 CDNA - SYNTHESIS 14
3.3.2 NESTED PCR FOR AMPLIFICATION OF IG GENE TRANSCRIPTS 15
3.4 IG GENE SEQUENCE ANALYSIS 16
3.5 DIRECTIONAL CLONING INTO EXPRESSION VECTORS 17
3.5.1 PREPARATION OF EUKARYOTIC EXPRESSION VECTORS 17
3.5.2 RESTRICTION ENDONUCLEASE DIGESTION 17
3.5.3 LIGATION 18
3.5.4 PREPARATION AND TRANSFORMATION OF COMPETENT BACTERIA 18
3.5.5 SCREENING BACTERIAL COLONIES BY PCR 19
3.5.6 PURIFICATION OF PLASMID DNA 20
3.6 RECOMBINANT ANTIBODY PRODUCTION 20
3.6.1 CELL CULTURE 20
3.6.2 PURIFICATION OF RECOMBINANT ANTIBODIES 21
3.7 ENZYME-LINKED IMMUNOSORBENT ASSAYS (ELISAS) 21
VI
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1011070308
DIGITALISIERT DURCH
IMAGE 2
TABLE OF CONTENTS
3.7.1 DETERMINATION OF RECOMBINANT IGG CONCENTRATIONS BY ELISA 21
3.7.2 DETERMINATION OF POLYREACTIVE ANTIBODIES BY ELISA 22
3.7.3 IMMUNOFLUORESCENCE ASSAYS (IFAS) 22
3.8 CHARACTERIZATION OF THE COGNATE ANTIGEN 23
3.8.1 SCREENING FOR AUTOANTIBODY SPECIFICITIES 23
3.8.2 PREPARATION OF CELL LYSATES 23
3.8.3 PRECLEAR AND IMMUNOPRECIPITATION 23
3.8.4 COOMASSIE STAINING 24
3.9 STATISTICAL ANALYSIS 24
4 RESULTS 25
4.1 CLINICAL FEATURES OF FOUR ANALYZED SSC PATIENTS 25
4.2 ISOLATION OF BLOOD IGG* MEMORY B CELLS AND BONE MARROW PLASMA CELLS
FROM SSC PATIENTS 26
4.3 IG H CHAIN GENE SEGMENT USAGE AND CDR3 FEATURES IN IGG* MEMORY B
CELLS
AND IGG* PLASMA CELLS FROM SSC PATIENTS 26
4.4 IG L CHAIN GENE SEGMENT USAGE IN IGG MEMORY B CELLS AND IGG F
PLASMA CELLS FROM SSC PATIENTS... 28
4.5 SOMATIC HYPERMUTATIONS AND IGG SUBCLASS DISTRIBUTION IN IGG* MEMORY
B CELLS
AND IGG* PLASMA CELLS IN SSC PATIENTS 30
4.6 SELF-REACTIVITY AMONG IGG* MEMORY B CELL AND IGG* PLASMA CELL
ANTIBODIES IN SSC PATIENTS 32
4.6.1 SIMILAR LEVELS OF SELF-REACTIVITY AMONG IGG* MEMORY B CELL
ANTIBODIES IN SSC PATIENTS
AND HEALTHY DONORS 33
4.6.2 INCREASED SELF-REACTIVITY AMONG IGG* PLASMA CELL ANTIBODIES IN SSC
PATIENTS 34
4.7 POLYREACTIVITY AMONG IGG* MEMORY B CELL AND IGG* PLASMA CELL
ANTIBODIES IN SSC PATIENTS 36
4.7.1 IGG* MEMORY B CELLS FROM SSC PATIENTS EXPRESS COMPARABLE LEVELS OF
POLYREACTIVE ANTIBODIES AS HEALTHY DONORS 36
4.7.2 IGG* PLASMA CELLS FROM SSC PATIENTS EXPRESS HIGHER LEVELS OF
POLYREACTIVE ANTIBODIES COMPARED TO HEALTHY DONORS 37
4.8 IGG* MEMORY B CELLS AND IGG* PLASMA CELLS EXPRESS DISEASE ASSOCIATED
ANTIBODIES IN SSC PATIENTS 39
4.8.1 IGG* MEMORY B CELL FROM SSC PATIENT EXPRESSES DISEASE ASSOCIATED
ANTI-SCL-70 ANTIBODY 39
4.8.2 IGG* PLASMA CELLS FROM SSC PATIENTS EXPRESS DISEASE ASSOCIATED
AUTOANTIBODIES 40
4.9 HIGH LEVELS OF SELF- AND POLYREACTIVE ANTIBODIES ARE EXPRESSED BY
IGG* PLASMA CELLS IN SSC 42
5 DISCUSSION 44
5.1 IG GENE REPERTOIRE IN SSC 44
5.2 ORIGIN OF SELF- AND POLYREACTIVITY IN SSC 45
5.3 SSC ASSOCIATED AUTOANTIBODIES 48
5.4 POSSIBLE THERAPEUTIC APPROACHES TO TARGET THE MEMORY OR PLASMA CELL
COMPARTMENT IN SSC 50
5.5 CONCLUSION AND OUTLOOK 51
6 REFERENCES 53
VII
IMAGE 3
TABLE OF CONTENTS
7 SUPPLEMENTAL MATERIAL 62
7.1 ANTIBODIES 62
7.2 ANTIGENS 62
7.3 BACTERIA 62
7.4 BACTERIAL CULTURE MEDIA 62
7.5 BUFFERS AND SOLUTIONS 63
7.6 CELL LINES 63
7.7 CELL CULTURE MEDIUM 64
7.8 CHEMICALS 64
7.9 COMMERCIAL KITS 64
7.10 PRECAST SDS-GELS 64
7.11 ENZYMES 64
7.12 EXPRESSION VECTORS 65
7.13 NUCLEOTIDES AND NUCLEIC ACIDS 65
7.14 REAGENTS 65
7.15 SOFTWARE 65
7.16 WEB RESOURCES 65
8 SUPPLEMENTAL FIGURES 66
8.1 THE HUMAN IG HEAVY AND LIGHT CHAIN LOCI 66
8.2 SINGLE CELL RT-PCR STRATEGY 67
8.3 EXPRESSION VECTOR MAPS 68
8.4 GATING STRATEGY FOR IGG* MEMORY B CELL ISOLATION 68
8.5 SEQUENCE ANALYSIS DATA FROM IGG* MEMORY B CELL AND IGG PLASMA CELL
ANTIBODIES FROM SSC PATIENTS 69
8.6 IMMUNOBLOT OF ANTIBODY SSC(PC)01-141 70
9 SUPPLEMENTAL TABLES 71
9.1 PRIMER FOR AMPLIFICATION OF HUMAN IG GENES 71
9.2 PRIMER FOR SEQUENCING AND BACTERIAL COLONY INSERT SCREENING 72
9.3 SEQUENCE DATA AND REACTIVITY OF IGG MEMORY B CELL AND IGG* PLASMA
CELL ANTIBODIES FROM SSC PATIENTS 73
10 APPENDIX 77
10.1 ACKNOWLEDGEMENTS 77
10.2 CURRICULUM VITAE 78
10.3 POSTERS AND PUBLICATIONS 80
10.4 SELBSTAENDIGKEITSERKLAERUNG 81
VIII
|
| any_adam_object | 1 |
| author | Zoller, Markus |
| author_facet | Zoller, Markus |
| author_role | aut |
| author_sort | Zoller, Markus |
| author_variant | m z mz |
| building | Verbundindex |
| bvnumber | BV037409754 |
| classification_rvk | YG 6093 |
| ctrlnum | (OCoLC)918319148 (DE-599)DNB1011070308 |
| dewey-full | 616.544 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 616 - Diseases |
| dewey-raw | 616.544 |
| dewey-search | 616.544 |
| dewey-sort | 3616.544 |
| dewey-tens | 610 - Medicine and health |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| isbn | 9783866649545 |
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| spelling | Zoller, Markus Verfasser aut IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis Markus Zoller Berlin Mensch & Buch Verl. 2011 81 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Berlin, Humboldt-Univ., Diss., 2011 Sklerodermie (DE-588)4181673-0 gnd rswk-swf Immunglobulin G (DE-588)4127637-1 gnd rswk-swf Autoantikörper (DE-588)4215429-7 gnd rswk-swf Plasmazelle (DE-588)4174842-6 gnd rswk-swf B-Lymphozyt (DE-588)4137121-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Sklerodermie (DE-588)4181673-0 s Autoantikörper (DE-588)4215429-7 s Immunglobulin G (DE-588)4127637-1 s B-Lymphozyt (DE-588)4137121-5 s Plasmazelle (DE-588)4174842-6 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022562248&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zoller, Markus IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis Sklerodermie (DE-588)4181673-0 gnd Immunglobulin G (DE-588)4127637-1 gnd Autoantikörper (DE-588)4215429-7 gnd Plasmazelle (DE-588)4174842-6 gnd B-Lymphozyt (DE-588)4137121-5 gnd |
| subject_GND | (DE-588)4181673-0 (DE-588)4127637-1 (DE-588)4215429-7 (DE-588)4174842-6 (DE-588)4137121-5 (DE-588)4113937-9 |
| title | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis |
| title_auth | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis |
| title_exact_search | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis |
| title_full | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis Markus Zoller |
| title_fullStr | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis Markus Zoller |
| title_full_unstemmed | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis Markus Zoller |
| title_short | IgG + memory B cell and IgG + plasma cell antibodies in patients with Systemic Sclerosis |
| title_sort | igg memory b cell and igg plasma cell antibodies in patients with systemic sclerosis |
| topic | Sklerodermie (DE-588)4181673-0 gnd Immunglobulin G (DE-588)4127637-1 gnd Autoantikörper (DE-588)4215429-7 gnd Plasmazelle (DE-588)4174842-6 gnd B-Lymphozyt (DE-588)4137121-5 gnd |
| topic_facet | Sklerodermie Immunglobulin G Autoantikörper Plasmazelle B-Lymphozyt Hochschulschrift |
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