Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice
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100 | 1 | |a Schneeweiß, Anna Maria Birthe |d 1991- |e Verfasser |0 (DE-588)1178714942 |4 aut | |
245 | 1 | 0 | |a Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |c vorgelegt von Anna Maria Birthe Schneeweiß, M. Sc. |
264 | 1 | |a Münster |c 2018 | |
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336 | |b txt |2 rdacontent | ||
337 | |b n |2 rdamedia | ||
338 | |b nc |2 rdacarrier | ||
502 | |b Dissertation |c Westfälische-Wilhelms-Universität Münster |d 2019 |g Beschränkt für den Austausch | ||
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999 | |a oai:aleph.bib-bvb.de:BVB01-031609979 |
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adam_text | TABLE OF CONTENTS
1
INTRODUCTION..............................................................................................................................
1
1.1
PSORIASIS..............................................................................................................................
1
1.1.1 CLINICAL
PICTURE.........................................................................................................2
1.1.2 SKIN PATHOLOGY AND HISTOLOGICAL HALLMARKS OF PSORIATIC PLAQUES
........................
2
1.1.3 GENETIC PREDISPOSITION & ENVIRONMENTAL FACTORS
.................................................
4
1.1.4 IMMUNOPATHOGENESIS O F PSORIASIS
..........................................................................
5
1.1.5 ROLE O F KERATINOCYTES FOR DISEASE INITIATION & PROGRESSION
..................................
8
1.1.6 ROLE O F NEUTROPHILS FOR DISEASE INITIATION & PROGRESSION
..................................
10
1.1.7 THERAPEUTIC
OPTIONS...............................................................................................
11
1.1.8 MOUSE MODELS O F
PSORIASIS....................................................................................14
1.1.8.1 IMIQUIMOD-INDUCED PSORIASIS-LIKE SKIN INFLAMMATION
...............................
14
1.2 BACTERIAL EFFECTOR
PROTEINS..............................................................................................
16
1.2.1 THE
YERSINIA
OUTER PROTEIN M
...............................................................................
16
1.3
OBJECTIVES........................................................................................................................
20
2 MATERIALS &
METHODS............................................................................................................
22
2.1
MATERIALS...........................................................................................................................22
2.1.1
EQUIPMENT................................................................................................................22
2.1.2
DISPOSABLES.............................................................................................................
23
2.1.3 PURCHASED CHEMICALS &
KITS..................................................................................
24
2.1.4 MEDIA &
BUFFERS.....................................................................................................
25
2.1.5 PRIMARY
ANTIBODIES.................................................................................................
27
2.1.6 SECONDARY
ANTIBODIES.............................................................................................
28
2.1.7
PRIMERS.........................................................................................................
28
2.1.8 MOUSE
LINES.............................................................................................................
28
2.1.9
SOFTWARE...................................................................................................................29
2.2
METHODS............................................................................................................................30
2.2.1 EXPERIMENTAL ANIMALS &
HOUSING.........................................................................30
2.2.2
IN VIVO
IMIQUIMOD-INDUCED PSORIASIS-LIKE SKIN INFLAMMATION MODEL
............
30
2.2.3 SUBCUTANEOUS TREATMENT WITH RYOPM AND ITS VARIANTS
.....................................
30
2.2.4 EPICUTANEOUS TREATMENT WITH
RYOPM...................................................................32
2.2.5 EVALUATION O F DISEASE
SEVERITY..............................................................................33
2.2.6 FLOW CYTOMETRIC ANALYSIS O F IMMUNE
CELLS.........................................................34
2.2.6.1 IN SKIN DRAINING LYMPH
NODES.........................................................................34
2.2.6.2 IN LESIONAL SK IN
....................................
.
........................................................
34
2.2.7 HISTOLOGICAL
EXAMINATION......................................................................................35
2.2.7.1 HEMATOXYLIN & EOSIN STAINING
......................................................................
35
2.2.7.2 IMMUNOFLUORESCENT
STAINING...........................................................................
35
2.2.7.3 TUNEL
STAINING..............................................................................................
35
2.2.7.4 FAM-FLICA STAINING O F ACTIVE
CASPASE-1...................................................36
2.2.8 RNA
EXTRACTION.......................................................................................................36
2.2.9 QUALITY CONTROL O F RN A
.........................................................................................36
2.2.10 CDNA SYNTHESIS AND QUANTITATIVE REAL-TIME PCR
...............................................
37
2.2.11 RNA-SEQ
ANALYSIS..................................................................................................
37
2.2.12 PROTEIN
ANALYSIS.......................................................................................................38
2.2.12.1 EXTRACTION O F TOTAL SKIN
PROTEIN.....................................................................38
2.2.12.2 TOTAL PROTEIN QUANTIFICATION BY BCA ASSAY
................................................
38
2.2.12.3 SDS-PAGE & WESTERN B LO T
........................................................................
39
2.2.12.4
LEGENDPLEX*.............................................................................................40
2.2.13 CELL CULTURE O F PRIMARY
CELLS..................................................................................41
2.2.13.1 ISOLATION OF PRIMARY NEUTROPHILS FROM BONE
MARROW.................................41
2.2.13.2 COATING O F CELL CULTURE PLATES FOR KERATINOCYTE CULTURE
..............................
42
2.2.13.3 ISOLATION OF PRIMARY KERATINOCYTES
................................................................
42
2.2.13.4 TREATMENT O F PRIMARY CELLS WITH RYOPM
.....................................................
42
2.2.13.5
CYTOSPINS.........................................................................................................
43
2.2.14 HUMAN SKIN
SAMPLES...............................................................................................43
2.2.15 STATISTICAL
ANALYSIS..................................................................................................
43
3
RESULTS....................................................................................................................................
44
3.1 SUBCUTANEOUSLY INJECTED RYOPM REDUCES CLINICAL SYMPTOMS O F
PSORIASIS-LIKE SKIN
INFLAMMATION IN M
ICE................................................................................................................
44
3.2 EPICUTANEOUS TREATMENT WITH
RYOPM..........................................................................
46
3.2.1 TOPICAL RYOPM TREATMENT INFLUENCES KERATINOCYTE ACTIVATION
..........................
51
3.2.2 TREATMENT WITH TOPICAL RYOPM HAS A MINOR EFFECT ON T C E LLS
.........................
53
3.2.3 TOPICALLY APPLIED RYOPM REDUCES NEUTROPHIL INFILTRATION IN
LESIONAL SKIN.... 55
3.3 TOPICALLY APPLIED RYOPM PENETRATES HUMAN SKIN
....................................................
59
3.4 THE FIRST THREE LRRS MEDIATE DISEASE AMELIORATION
..................................................
60
3.4.1 INFLUENCE OF RYOPM ON
KERATINOCYTES...................................................................64
3.4.2 MARGINAL ROLE O F T CELLS DURING RYOPM
TREATMENT.............................................69
3.4.3 REDUCED NUMBERS O F NEUTROPHILS IN LESIONAL SKIN IN WT MICE
........................
75
3.4.4 RNA-SEQ
ANALYSIS..................................................................................................
79
4
DISCUSSION...............................................................................................................................
93
4.1 RECOMBINANT YOPM AND YOPMUUU
-3
SIMILARLY AMELIORATE ONGOING SKIN-
INFLAMMATION WHEN INJECTED SUBCUTANEOUSLY
.........................................................................
93
4.1.1 POTENTIAL MECHANISM O F THE ANTI-INFLAMMATORY CAPACITY
..................................
99
4.1.2 ROLE O F T CELLS DURING RY OPM
TREATMENT...........................................................102
4.1.3 EFFECT O F RY OPM ON KERATINOCYTES
.....................................................................
104
4.1.4 EFFECT O F RYOPM ON
NEUTROPHILS.........................................................................109
4.2 POSSIBLE TRANSFER TO THE HUMAN
PATIENT......................................................................
111
5 CONCLUSION &
OUTLOOK.........................................................................................................117
|
any_adam_object | 1 |
author | Schneeweiß, Anna Maria Birthe 1991- |
author_GND | (DE-588)1178714942 |
author_facet | Schneeweiß, Anna Maria Birthe 1991- |
author_role | aut |
author_sort | Schneeweiß, Anna Maria Birthe 1991- |
author_variant | a m b s amb ambs |
building | Verbundindex |
bvnumber | BV046231490 |
ctrlnum | (OCoLC)1126564690 (DE-599)DNB1178343456 |
discipline | Medizin |
format | Thesis Book |
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physical | 119, XXXVII Blätter Illustrationen 30 cm 1 CD-ROM |
publishDate | 2018 |
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record_format | marc |
spelling | Schneeweiß, Anna Maria Birthe 1991- Verfasser (DE-588)1178714942 aut Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice vorgelegt von Anna Maria Birthe Schneeweiß, M. Sc. Münster 2018 119, XXXVII Blätter Illustrationen 30 cm 1 CD-ROM txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische-Wilhelms-Universität Münster 2019 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1178343456/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031609979&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
spellingShingle | Schneeweiß, Anna Maria Birthe 1991- Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |
subject_GND | (DE-588)4113937-9 |
title | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |
title_auth | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |
title_exact_search | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |
title_full | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice vorgelegt von Anna Maria Birthe Schneeweiß, M. Sc. |
title_fullStr | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice vorgelegt von Anna Maria Birthe Schneeweiß, M. Sc. |
title_full_unstemmed | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice vorgelegt von Anna Maria Birthe Schneeweiß, M. Sc. |
title_short | Truncated rYopM reduces keratinocyte activation and neutrophil infiltration in ongoing IMQ-induced psoriasiform dermatitis in mice |
title_sort | truncated ryopm reduces keratinocyte activation and neutrophil infiltration in ongoing imq induced psoriasiform dermatitis in mice |
topic_facet | Hochschulschrift |
url | http://d-nb.info/1178343456/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031609979&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
work_keys_str_mv | AT schneeweißannamariabirthe truncatedryopmreduceskeratinocyteactivationandneutrophilinfiltrationinongoingimqinducedpsoriasiformdermatitisinmice |