The role of microRNA-155 in innate immunity and immunopathology
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV040660547 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t | ||
| 008 | 130109s2012 ad|| m||| 00||| eng d | ||
| 035 | |a (OCoLC)826576185 | ||
| 035 | |a (DE-599)BSZ37602481X | ||
| 040 | |a DE-604 |b ger | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-355 | ||
| 100 | 1 | |a Hedlund, Sebastian |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a The role of microRNA-155 in innate immunity and immunopathology |c Sebastian Hedlund |
| 264 | 1 | |c 2012 | |
| 300 | |a XI, 124 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Freiburg i. Br., Univ., Diss., 2012 | ||
| 650 | 0 | 7 | |a Immunpathologie |0 (DE-588)4014133-0 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a miRNS |0 (DE-588)7736808-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Immunreaktion |0 (DE-588)4133841-8 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Knockout |g Molekulargenetik |0 (DE-588)4671699-3 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Immunreaktion |0 (DE-588)4133841-8 |D s |
| 689 | 0 | 1 | |a Immunpathologie |0 (DE-588)4014133-0 |D s |
| 689 | 0 | 2 | |a miRNS |0 (DE-588)7736808-3 |D s |
| 689 | 0 | 3 | |a Knockout |g Molekulargenetik |0 (DE-588)4671699-3 |D s |
| 689 | 0 | |5 DE-604 | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025487313&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025487313 | ||
Datensatz im Suchindex
| _version_ | 1804149783535812608 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
ERKLARUNGEN I
ACKNOWLEDGEMENTS N
SUMMARY IN
ABBREVIATIONS IV
TABLE OF CONTENTS VIII
INTRODUCTION 1
1.1 IMMUNITY 2
1.1.2 INNATE IMMUNITY..... 2
1.1.2.1 ANTIGEN-PRESENTING CELLS 3
1.1.2.2 TOLL-LIKE RECEPTORS 4
1.1.2.3 DENDRITIC CELLS 6
1.1.2.4 MACROPHAGES 7
1.1.3 ADAPTIVE IMMUNITY 8
1.2 IMMUNE RESPONSE 9
1.2.1 IMMUNE RESPONSE 9
1.2.2 CONTACT HYPERSENSITIVITY 10
1.3 NON-CODING R N A 1 1
1.4 MICRORNA 1 3
1.4.1 HISTORY 13
1.4.2 FUNCTION 14
1.4.3 BIOGENESIS 15
1.4.4 MICRORNA IN DISEASE AND INFLAMMATION 17
1.4.5 HEMATOPOIETIC REGULATION BY MICRORNAS 18
1.5 MICRORNA-1 5 5 2 0
1.5R1-MIR-155 REGULATION 21
1.5.2 MIR-155 RELATED MALIGNANCIES 22
1.5.2.1 HEMATOPOIESIS 22
1.5.2.2 CANCER 22
1.5.2.2.2 LYMPHOMA 22
1.5.2.2.2 LEUKEMIA 23
1.5.2.2.3 TUMORS ....23
1.5.2.3 CARDIOVASCULAR DISEASE 23
1.5.2.4 ACUTE GRAFT-VERSUS-HOST DISEASE (AGVHD) 24
1.5.2.5 AUTOIMMUNE DISEASE 24
1.5.2.5.1 MULTIPLE SCLEROSIS 24
1.5.2.5.2 RHEUMATOID ARTHRITIS 25
1.5.2.6 ASTHMA 25
1.5.3 MIR-155 IN IMMUNITY 25
1.6 AIM 2 7
VIII
HTTP://D-NB.INFO/1029248168
IMAGE 2
TABLE O F CONTENTS
MATERIALS 2 8
3.1 LABORATORY EQUIPMENT 29
3.2 LAB WARE 29
3.3 LABORATORY REAGENTS, KITS, CHEMICALS AND ENZYMES 30 3.4 BUFFERS 32
3.5 ACRYLAMIDE GELS 33
3.5.1 SEPARATING GEL 33
3.5.2 STACKING GEL 34
3.6 ANTIBODIES 34
3.7 PRIMERS & PROBES 35
METHODS 3 7
4.1 MOUSE STRAINS 3 8
4.1.1 MICE EUTHANIZATION 38
4 . 2 CELL BIOLOGY 3 9
4.2.1 CELL ISOLATION 39
4.2.1.2 ERYTHROCYTE LYSIS 39
4.2.2 CELL CULTURE 41
4.2.3 CELL LINES 41
4.2.4 PRIMARY CELL CULTURE 42
4.2.4.3.1 M1-MACROPHAGE DIFFERENTIATION 43
4.2.4.3.2 M2-MACROPHAGE DIFFERENTIATION 43
4 . 3 IMMUNOLOGY 4 5
4.3.4 TRANSLATIONAL REPRESSION OF MYELOID CELLS 46
4.3.5 ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) 47
4.3.6 GRIESS TEST 47
4.3.8 INDUCTION OF CONTACT HYPERSENSITIVITY (CHS) 48
4.3.8.1 IMMUNIZATION AND INDUCTION OF CHS 48
4.3.8.2 ADOPTIVE DC TRANSFER AND INDUCTION OF CHS 48
4 . 4 MOLECULAR BIOLOGY 5 0
4.4.1 GENOTYPING OF MIR-155 KNOCKOUT MOUSE 50
4.4.2 RNA EXTRACTION 50
4.4.2.1 MIRNEASYKIT 50
4.4.2.2 TRIZOL METHOD 51
4.4.3 REVERSE TRANSCRIPTASE PCR (RT-PCR) 52
4.4.4 REAL-TIME PCR (QPCR) 53
4.4.5 MICROARRAY 55
4.4.6 NORTHERN BLOT 56
4.4.7 WESTERN BLOT BY SDS-PAGE 57
4.4.7.1 TOTAL PROTEIN QUANTIFICATION 58
4.4.8 PROLIFERATION ASSAY 59
IX
IMAGE 3
TABLE O F CONTENTS
RESULTS 6 0
5.1 CELL CULTURE 6 1
5.1.1 OBTAINING DC AND M T FROM BM CELLS BY PRIMARY CELL CULTURE 61
5.1.2 CONVENTIONAL DC DERIVES FROM THE COMMON DENDRITIC PRECURSOR 62 5.2
MIR-155 EXPRESSION 6 4
5.2.1 TISSUE EXPRESSION OF MIR-155 IS LOW UNDER STEADY-STATE CONDITIONS
64
5.2.2 MIR-155 IS CORRELATIVELY INDUCED UPON IMMUNOLOGICAL ACTIVATION 65
5.2.2.1 FLT3 CULTURED CELLS 65
5.2.2.2 GM-CSF CULTURED CELLS 65
5.2.2.3 M-CSF CULTURED CELLS 65
5.2.2.4 TLR-ACTIVATION OF B-CELLS INDUCE MIR-155 EXPRESSION 66 5.2.2.5
EXPRESSION OF PU1 CORRELATES WITH MIR-155 EXPRESSION 66 5.2.3 EXPRESSION
LEVEL OF MIR-155 REACHES A MAXIMUM AFTER 8-10 H OF STIMULATION AND
REMAINS FOR 48 H 68
5.2.4 MIR-155 CAN BE INDUCED BY TLR-LIGANDS BUT REQUIRES DE NOVO
PROTEINS 69
5.2.5 MIR-155 EXPRESSION IN PERITONITIS 70
5.3 THE MIR-1 5 5 KNOCKOUT MOUSE 7 2
5.3.1 VERIFICATION OF MIR-155 KNOCKOUT MOUSE GENOTYPE 72 5.3.2 MIR-155
KNOCKOUT MICE HAVE A FUNCTIONAL REPRODUCTION 73 5.4 MIR-1 55-DEFICIENT
MACROPHAGES 74
5.4.1 MYELOID COMPOSITION OF CELLULAR COMPARTMENTS IN MIR-155 KO MICE 74
5.4.2 M2-PERITONEAL MACROPHAGES IN STEADY-STATE CONDITIONS 76 5.4.3
MIR-155 IS NOT REQUIRED FOR MACROPHAGE DIFFERENTIATION 77 5.4.4
MIR-155-DEFICIENT MACROPHAGES SHOW NORMAL ACTIVATION STATUS 79 5.4.5
MIR-155 EXPRESSION IS INVOLVED IN ALL PARTS OF THE INFLAMMATORY
RESPONSE IN MACROPHAGES 80
5.5 MIR-1 55-DEFICIENT DC 82
5.5.1 MIR-155 KO MICE ARE RESISTANT TO CONTACT HYPERSENSITIVITY 82 5.5.2
SUPPRESSION OF CHS IN MIR-155 KO MICE IS DUE TO DC DEFECT 83 5.5.3
MEMBRANE-ASSOCIATED GENES ARE DYSREGULATED IN MIR-155DEFICIENT DC 84
5.5.4 FUNCTIONAL IN VITRO MIR-155 KO DC ACTIVATION 86
5.5.5 IN VIVO DC MIGRATION IS FUNCTIONAL IN MIR-155 KO MICE 88 5.5.6 T
CELL ACTIVATING POTENTIAL OF MIR-155-DEFICIENT DC 90 5.5.7 MIR-155
EXPRESSION IS REQUIRED IN DC TO ENABLE THEM TO INDUCE PROPER T CELL
PROLIFERATION 92
X
IMAGE 4
TABLE O F CONTENTS
DISCUSSION 9 3
6.1 D C AND M O CELL CULTURE 9 5
6 . 2 MLR-155 EXPRESSION CORRELATES WITH THE EXPRESSION OF
INFLAMMATION MARKERS 9 6
6.3 MLR-155 EXPRESSION CAN BE INDUCED BY SEVERAL TLR LLGANDS AND
REQUIRES D E N O V O PROTEINS 9 7
6 . 4 PERITONITIS ONLY INDUCES A MILD UPREGULATION OF MIR-1 5 5 9 8
6 . 5 M I R - 1 5 5 KNOCKOUT MOUSE VERIFICATION AND REPRODUCTION 9 9 6 .
6 M 2 MACROPHAGES RESIDE IN THE PERITONEAL CAVITY IN STEADY-STATE
CONDITIONS, BUT ARE OVERTAKEN BY M 1 MACROPHAGES UPON INFLAMMATION 9 9
6 . 7 MLR-155 IS NOT ACTIVELY REGULATING MACROPHAGE
DIFFERENTIATION 1 0 1
6 . 8 M I R - 1 5 5 K O MICE FAIL TO INDUCE A CONTACT HYPERSENSITIVITY
RESPONSE DUE TO IMPAIRED D C FUNCTION 1 0 3
6 . 9 CONCLUDING REMARKS 1 0 6
REFERENCES 1 0 7
APPENDIX 1 2 1
APPENDIX A - M I R - 1 5 5 K O MOUSE GENOTYPE 1 2 1
APPENDIX B - EXPRESSION LEVEL OF PU1 IS AFFECTED BY MIR-1 55 EXPRESSION
1 2 3
APPENDIX C - MIR-1 5 5 K O B CELLS 1 2 4
XI
|
| any_adam_object | 1 |
| author | Hedlund, Sebastian |
| author_facet | Hedlund, Sebastian |
| author_role | aut |
| author_sort | Hedlund, Sebastian |
| author_variant | s h sh |
| building | Verbundindex |
| bvnumber | BV040660547 |
| ctrlnum | (OCoLC)826576185 (DE-599)BSZ37602481X |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01537nam a2200385 c 4500</leader><controlfield tag="001">BV040660547</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">130109s2012 ad|| m||| 00||| eng d</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)826576185</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BSZ37602481X</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Hedlund, Sebastian</subfield><subfield code="e">Verfasser</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">The role of microRNA-155 in innate immunity and immunopathology</subfield><subfield code="c">Sebastian Hedlund</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2012</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">XI, 124 Bl.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">Freiburg i. Br., Univ., Diss., 2012</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Immunpathologie</subfield><subfield code="0">(DE-588)4014133-0</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">miRNS</subfield><subfield code="0">(DE-588)7736808-3</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Immunreaktion</subfield><subfield code="0">(DE-588)4133841-8</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Knockout</subfield><subfield code="g">Molekulargenetik</subfield><subfield code="0">(DE-588)4671699-3</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="689" ind1="0" ind2="0"><subfield code="a">Immunreaktion</subfield><subfield code="0">(DE-588)4133841-8</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="1"><subfield code="a">Immunpathologie</subfield><subfield code="0">(DE-588)4014133-0</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="2"><subfield code="a">miRNS</subfield><subfield code="0">(DE-588)7736808-3</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="3"><subfield code="a">Knockout</subfield><subfield code="g">Molekulargenetik</subfield><subfield code="0">(DE-588)4671699-3</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2=" "><subfield code="5">DE-604</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025487313&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-025487313</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV040660547 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:28:31Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025487313 |
| oclc_num | 826576185 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | XI, 124 Bl. Ill., graph. Darst. |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
| record_format | marc |
| spelling | Hedlund, Sebastian Verfasser aut The role of microRNA-155 in innate immunity and immunopathology Sebastian Hedlund 2012 XI, 124 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Freiburg i. Br., Univ., Diss., 2012 Immunpathologie (DE-588)4014133-0 gnd rswk-swf miRNS (DE-588)7736808-3 gnd rswk-swf Immunreaktion (DE-588)4133841-8 gnd rswk-swf Knockout Molekulargenetik (DE-588)4671699-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Immunreaktion (DE-588)4133841-8 s Immunpathologie (DE-588)4014133-0 s miRNS (DE-588)7736808-3 s Knockout Molekulargenetik (DE-588)4671699-3 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025487313&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hedlund, Sebastian The role of microRNA-155 in innate immunity and immunopathology Immunpathologie (DE-588)4014133-0 gnd miRNS (DE-588)7736808-3 gnd Immunreaktion (DE-588)4133841-8 gnd Knockout Molekulargenetik (DE-588)4671699-3 gnd |
| subject_GND | (DE-588)4014133-0 (DE-588)7736808-3 (DE-588)4133841-8 (DE-588)4671699-3 (DE-588)4113937-9 |
| title | The role of microRNA-155 in innate immunity and immunopathology |
| title_auth | The role of microRNA-155 in innate immunity and immunopathology |
| title_exact_search | The role of microRNA-155 in innate immunity and immunopathology |
| title_full | The role of microRNA-155 in innate immunity and immunopathology Sebastian Hedlund |
| title_fullStr | The role of microRNA-155 in innate immunity and immunopathology Sebastian Hedlund |
| title_full_unstemmed | The role of microRNA-155 in innate immunity and immunopathology Sebastian Hedlund |
| title_short | The role of microRNA-155 in innate immunity and immunopathology |
| title_sort | the role of microrna 155 in innate immunity and immunopathology |
| topic | Immunpathologie (DE-588)4014133-0 gnd miRNS (DE-588)7736808-3 gnd Immunreaktion (DE-588)4133841-8 gnd Knockout Molekulargenetik (DE-588)4671699-3 gnd |
| topic_facet | Immunpathologie miRNS Immunreaktion Knockout Molekulargenetik Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025487313&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT hedlundsebastian theroleofmicrorna155ininnateimmunityandimmunopathology |