Autoimmunity in complex regional pain syndrome:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Giessen
VVB Laufersweiler
2011
|
| Ausgabe: | 1. Aufl. |
| Schriftenreihe: | Edition scientifique
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | VIII, 112 S. Ill., graph. Darst. |
| ISBN: | 9783835958258 |
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Datensatz im Suchindex
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| adam_text | IMAGE 1
INDEX
INDEX I
LIST OF ABBREVIATIONS IV
LIST OF TABLES VI
LIST OF FIGURES VII
I - INTRODUCTION 1
1.1. COMPLEX REGIONAL PAIN SYNDROME 1
1.1.1. INCIDENCE 1
1.1.2. ETIOLOGY 2
1.1.3. PATHOGENESIS AND MECHANISMS 3
1.1.3.1. INFLAMMATION, NEUROGENIC INFLAMMATION AND HYPOXIC CHANGES 3
1.1.3.2. AUTONOMIE NERVOUS SYSTEM DYSREGULATION 5
1.1.3.3. SENSITIZATION AND CORTICAL REORGANIZATION 7
,1.1.3.4. POSSIBLE INVOLVEMENT OF THE IMMUNE SYSTEM 8
1.1.4. CLINICAL FEATURES 9
1.1.5. DIAGNOSIS 12
1.1.6. TREATMENT 14
1.2. AUTOIMMUNITY 14
1.2.1. MECHANISM OF AUTOIMMUNITY 16
1.2.1.1. MHC CONTROL AND GENETIC BACKGROUND 16
1.2.1.2. DISRUPTION IN THE LEVEL OR ACTIVITY OF REGULATORY PROTEINS AND
CELLS17 1.2.1.3. ALTERED PROTEINS 17
1.2.1.4. APOPTOSIS AND AUTOIMMUNITY TO SEQUESTERED PROTEINS 18 1.2.1.5.
MOLECULAR MIMICRY 19
1.2.1.6. NATURAL ANTIBODIES AND AUTOANTIBODIES 19
II-AIM 21
III - MATERIALS AND METHODS 22
3.1. PATIENTS 22
3.2. MATERIALS 22
3.2.1. CHEMICALS AND SOLUTIONS 22
3.2.2. CONSUMABLES 24
3.2.3. INSTRUMENTS 25
3.2.4. MOLECULAR BIOLOGY AND BIOCHEMISTRY KITS 26
3.2.5. BUFFERS AND SOLUTIONS 26
3.2.6. MEDIA AND SOLUTIONS FOR CELL CULTURE 27
3.2.7. CELL LINES 28
3.2.8. ANTIBODIES 28
3.2.9. PRIMERS 29
I
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1016488181
DIGITALISIERT DURCH
IMAGE 2
INDEX
3.2.10. SOFTWARE 29
3.3. METHODS 30
3.3.1. EXPERIMENT SET 1: AUTOANTIBODY DETECTION BY FLOW CYTOMETRY ON THE
SURFACE OF DIFFERENTIATED AUTONOMIE CELLS 30
3.3.1.1. CELL CULTURE 30
3.3.1.2. DIFFERENTIATION OF NEUROBLASTOMA CELLS 30
3.3.1.3. ISOLATION OF SYMPATHETIC NEURONS 31
3.3.1.4. ISOLATION OF GUT MYENTERIC PLEXUS 31
3.3.1.5. AUTOANTIBODY DETECTION BY FLOW CYTOMETRY 32
3.3.1.6. IDENTIFICATION OF NEURONAL CELLS 32
3.3.1.7. PCR EXPERIMENTS 33
3.3.2. EXPERIMENT SET 2: IDENTIFICATION OF PUTATIVE AUTOANTIGENS
TARGETED BY CRPS SERUM ANTIBODIES 34
3.3.2.1. CELL CULTURE 34
3.3.2.2. AFFINITY CHROMATOGRAPHY 34
3.3.2.3. PREPARATION OF CARDIOMYOCYTE CULTURES 34
3.3.2.4. PEPTIDES 35
3.3.2.5. PHARMACOLOGICAL ASSAYS 35
3.3.2.6. ENZYME IMMUNOASSAY 36
3.3.3. EXPERIMENT SET 3: DETERMINATION OF IMMUNOGLOBULIN SUBCLASSES OF
CRPS PATHOGENIC ANTIBODIES 37
3.3.4. EXPERIMENT SET 4: CRPS PATHOGENIC ANTIBODIES BIND TO THE SURFACE
OF STABLE TRANSFECTED CELLS 37
3.3.4.1. ANTIBODY BINDING MEASURED BY FLOW CYTOMETRY 37
3.3.4.2. PCR EXPERIMENTS 38
3.3.5. EXPERIMENT SET 5: FUNCTIONAL ASSAYS 38
3.3.5.1. FUNCTIONAL CARDIOMYOCYTE ASSAY 38
3.3.5.2. RECEPTOR EXPRESSION 39
3.3.5.3. CALCIUM IMAGING 39
3.3.6. EXPERIMENT SET 6: CYTOTOXICITY OF PATHOGENIC AUTOANTIBODIES 40
3.3.6.1. LDH 40
3.3.6.2. WST-1 ASSAY 40
3.3.7. STATISTICAL ANALYSIS 41
IV - RESULTS 42
4.1. EXPERIMENT SET 1: SERUM AUTOANTIBODIES IN COMPLEX REGIONAL PAIN
SYNDROME BIND TO A DIFFERENTIATION-DEPENDENT NEURONAL
SURFACE-AUTOANTIGEN. 42 4.1.1. PATIENTS 42
4.1.2. SURFACE BINDING TO PRIMARY CULTURED AUTONOMIE NEURONS 43 4.1.3.
BINDING TO UNDIFFERENTIATED SH-SY5Y NEUROBLASTOMA CELLS 45 4.1.4.
BINDING TO DIFFERENTIATED SH-SY5Y NEUROBLASTOMA CELLS 46
4.1.5. ASSOCIATION WITH CLINICAL DATA ; 49
4.2. EXPERIMENT SET 2: CRPS IMMUNOGLOBULINS BIND TO THE SURFACE OF
CARDIOMYOCYTES ON THE M2R AND SS2AR 50
II
IMAGE 3
INDEX
4.2.1. PATIENTS 50
4.2.2 - CRPS IMMUNOGLOBULINS EXHIBIT AGONISTIC EFFECTS ON THE M2R AND
THE SS2AR IN SPONTANEOUSLY BEATING CARDIOMYOCYTES 51
4.2.3. THE IGG EFFECT OF CRPS CAN BE BLOCKED BY SPECIFIC PEPTIDES
DERIVED FROM THE SECOND EXTRACELLULAR LOOP OF THE M2R AND THE SS2AR 56
4.2.4. PEPTIDE CHARACTERIZATION BY ELISA 59
4.2.5. RECEPTOR AUTOANTIBODIES IN CRPS DO NOT CROSS-REACT BETWEEN M2R
ANDSS2AR 61
4.3. EXPERIMENT SET 3: IMMUNOGLOBULINS SUBCLASSES OF PATHOGENIC
ANTIBODIES 63
4.3.1. IGG SUBCLASSES CHARACTERIZATION OF CRPS ANTIBODIES 63 4.4.
EXPERIMENT SET 4: CRPS PATHOGENIC ANTIBODIES BIND TO THE SURFACE OF
HM2R- AND HSS2AR-TRANSFECTED CHO CELLS 63
4.5. EXPERIMENT SET 5: FUNCTIONAL ASSAYS 67
4.5.1. PATHOGENIC ANTIBODIES SHOW AGONISTIC-FUNCTIONAL EFFECTS BUT DO
NOT INDUCE DESENSITIZATION 67
4.5.2. SS2AR AND M2R AUTOANTIBODIES DO NOT DOWNREGULATE THEIR
CORRESPONDING RECEPTORS ON THE CELL SURFACE 70
4.5.3. SS2AR AND M2R AUTOANTIBODIES CAUSE RECEPTOR DEPENDENT INCREASE IN
CYTOSOLIC CA 2+ CONCENTRATION IN TRANSFECTED CELL LINES 71
4.5.4. ASSOCIATION WITH CLINICAL DATA 75
4.6. EXPERIMENT SET 6: CRPS ANTIBODIES DO NOT INDUCE CYTOTOXICITY 75
V - DISCUSSION 78
VI - SUMMARY 92
VII - ZUSAMMENFASSUNG 94
VIII - REFERENCES 96
IX - ACKNOWLEDGMENTS 107
X - PUBLICATIONS 108
XI-APPENDIX 109
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| isbn | 9783835958258 |
| language | English |
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| spelling | Kohr, Danielle Verfasser aut Autoimmunity in complex regional pain syndrome by Danielle Kohr 1. Aufl. Giessen VVB Laufersweiler 2011 VIII, 112 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Edition scientifique Zugl.: Gießen, Univ., Diss., 2011 Vegetatives Nervensystem (DE-588)4062444-4 gnd rswk-swf Autoimmunität (DE-588)4319435-7 gnd rswk-swf Komplexes regionales Schmerzsyndrom (DE-588)1059232081 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Komplexes regionales Schmerzsyndrom (DE-588)1059232081 s Vegetatives Nervensystem (DE-588)4062444-4 s Autoimmunität (DE-588)4319435-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024708151&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Kohr, Danielle Autoimmunity in complex regional pain syndrome Vegetatives Nervensystem (DE-588)4062444-4 gnd Autoimmunität (DE-588)4319435-7 gnd Komplexes regionales Schmerzsyndrom (DE-588)1059232081 gnd |
| subject_GND | (DE-588)4062444-4 (DE-588)4319435-7 (DE-588)1059232081 (DE-588)4113937-9 |
| title | Autoimmunity in complex regional pain syndrome |
| title_auth | Autoimmunity in complex regional pain syndrome |
| title_exact_search | Autoimmunity in complex regional pain syndrome |
| title_full | Autoimmunity in complex regional pain syndrome by Danielle Kohr |
| title_fullStr | Autoimmunity in complex regional pain syndrome by Danielle Kohr |
| title_full_unstemmed | Autoimmunity in complex regional pain syndrome by Danielle Kohr |
| title_short | Autoimmunity in complex regional pain syndrome |
| title_sort | autoimmunity in complex regional pain syndrome |
| topic | Vegetatives Nervensystem (DE-588)4062444-4 gnd Autoimmunität (DE-588)4319435-7 gnd Komplexes regionales Schmerzsyndrom (DE-588)1059232081 gnd |
| topic_facet | Vegetatives Nervensystem Autoimmunität Komplexes regionales Schmerzsyndrom Hochschulschrift |
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