Holdings: The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall

The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall:

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Main Author:	Schäfer, Kerstin 1989- (Author)
Format:	Thesis Book
Language:	English
Published:	[Münster] 2018
Subjects:	Hochschulschrift
Online Access:	Inhaltsverzeichnis Inhaltsverzeichnis
Physical Description:	IV, 125 Seiten Illustrationen 30 cm

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MARC


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adam_text	CONTENTS ABSTRACT IV 1. INTRODUCTION 1 1.1. IM M U N ITY ......................................................................................................................... 1 1.1.1. BLOOD V ESSELS...................................................................................................... 2 1.1.2. N EU TRO P H ILS......................................................................................................... 7 1.2. LEUKOCYTE EXTRAVASATION CASCADE.................................................................................. 7 1.2.1. CAPTURING AND R O LLIN G ....................................................................................... 8 1.2.2. ACTIVATION AND A R R E S T ....................................................................................... 9 1.2.3. FIRM ADHESION AND C RA W LIN G .......................................................................... 9 1.2.4. T RANSM IGRATION................................................................................................... 10 1.3. INTEGRINS ON L E U K O C Y TE S................................................................................................ 12 1.3.1. INSIDE-OUT S IG N A LIN G .......................................................................................... 14 1.3.2. OUTSIDE-IN S IG N A LIN G .......................................................................................... 14 1.4. PLATELET ENDOTHELIAL CELL ADHESION MOLECULE-1 (PECAM-1) .................................... 15 1.5. ICAM-1 AND ICAM-2 ................................................................................................... 16 1.6. CD99 ............................................................................................................................... 17 1.7. C D 9 9 L 2 ............................................................................................................................ 21 1.8. PAIRED IMMUNE R E C E P TO RS............................................................................................. 22 1.9. EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS........................................................... 23 2. AIM OF THE STUDY 25 3. MATERIAL 26 3.1. C E LLS .................................................................................................................................. 26 3.2. A N IM A LS ............................................................................................................................ 26 3.3. A NTIBODIES......................................................................................................................... 27 3.4. P RIM E R............................................................................................................................... 28 3.5. ENZYM ES............................................................................................................................ 29 3.6. CHEMICALS AND R E A G E N TS................................................................................................ 29 3.7. BUFFERS AND M E D I A ......................................................................................................... 31 3.8. CONSUMABLE SUPPLIES...................................................................................................... 32 3.9. EQUIPMENT ..................................................................................................................... 33 4. METHODS 35 4.1. MOLECULAR BIOLOGICAL M E TH O D S....................................................................................... 35 4.1.1. MOUSE EAR LYSIS FOR G E N O TY P IN G .................................................................... 35 4.1.2. POLYMERASE CHAIN R E A C TIO N ............................................................................... 35 4.1.3. AGAROSE G E L......................................................................................................... 38 4.2. IN VIVO M E TH O D S ............................................................................................................ 38 4.2.1. WHOLE BLOOD CELL C O U N TS .................................................................................. 38 4.2.2. IRRADIATION AND BONE MARROW TRANSPLANTATION OF M IC E ............................... 39 4.2.3. WHOLE MOUNT STAINING OF MOUSE CREMASTER M USCLE ..................................... 39 4.2.4. ANALYSIS OF NEUTROPHIL EXTRAVASATION.............................................................. 39 4.2.5. INTRAVITAL MICROSCOPY OF THE CREMASTER MUSCLE USING AN INTRAVITAL UPRIGHT M ICROSCOPE......................................................................................................... 40 4.2.6. ANALYSIS OF LEUKOCYTE ROLLING, ADHESION AND TRANSMIGRATION USING AN UPRIGHT MICROSCOPE .......................................................................................... 41 4.2.7. INTRAVITAL MICROSCOPY OF THE CREMASTER MUSCLE USING A CONFOCAL LSM 880 MICROSCOPE WITH FAST AIRY S C A N .............................................. 41 4.2.8. ANALYSIS OF THE LEUKOCYTE TRANSMIGRATION P R O C E S S ..................................... 42 4.2.9. INDUCTION OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) .... 43 4.2.10. STAINING OF MOUSE BRAIN S ECTIONS.................................................................... 44 4.3. CELL C U LTU RE ..................................................................................................................... 44 4.3.1. GENERAL M E TH O D S................................................................................................ 44 4.3.2. CULTIVATION OF DIFFERENT CELL T Y P E S ................................................................. 45 4.3.3. PASSAGING OF CELLS ............................................................................................. 45 4.3.4. COUNTING OF C E LLS................................................................................................ 45 4.4. METHODS OF CELL B IO LO G Y ................................................................................................ 46 4.4.1. ISOLATION OF MURINE POLYMORPHONUCLEAR CELLS (PMNS) ............................... 46 4.4.2. ISOLATION OF PRIMARY MURINE LUNG MICROVASCULAR ENDOTHELIAL CELLS .... 46 4.4.3. MAGNETIC BEAD SORTING OF PRIMARY ENDOTHELIAL C E LLS .................................. 47 4.5. BIOCHEMICAL M ETHODS...................................................................................................... 47 4.5.1. COUPLING OF A-PECAM-1 ANTIBODIES TO D YNABEADS ..................................... 47 4.5.2. S D S -P A G E ......................................................................................................... 48 4.5.3. WESTERN B L O T ...................................................................................................... 48 4.6. STATISTICS ........................................................................................................................ 50 5. RESULTS 51 5.1. CD99 AND CD99L2 GENE INACTIVATION AFFECTS NEUTROPHIL EXTRAVASATION .............. 51 5.2. NEUTROPHILS OF CD99L2- / * MICE GET TRAPPED IN LAMININ A 5 LOW EXPRESSION R E G IO N S ........................................................................................................................... 54 5.3. GENE DEFICIENCY OF CD99 SHOWS DECREASED ADHESION AND TRANSMIGRATION AND KNOCKOUT OF CD99L2 RESULTS IN DECREASED TRANSMIGRATION ...................................... 57 5.4. OPTIMIZATION OF INTRAVITAL MICROSCOPY USING A CONFOCAL MICROSCOPE (ZEISS LSM 880) 59 5.5. GENE ABLATION OF CD99L2 RESULTS IN DECREASED SPEED OF ABLUMINAL CRAWLING IN V I V O .................................................................................................................................. 61 5.6. GENE DEFICIENCY OF CD99 AFFECTS ADHESION AND SPEED OF ABLUMINAL CRAWLING IN V I V O .................................................................................................................................. 70 5.7. ICAM-1 BLOCKING IN IL -LSS INJECTED MICE AFFECTS THE ABLUMINAL CRAWLING STEP . 77 5.8. THE ROLE OF CD99L2 IN TRANSMIGRATION OF LEUKOCYTES IN EAE-INJECTED MICE . . . 78 6. DISCUSSION 81 6.1. STUDYING CD99 AND CD99L2 FUNCTION USING ANTIBODIES AND GENE-DEFICIENT MICE 82 6.2. ROLE OF CD99 AND CD99L2 IN THE TRANSENDOTHELIAL MIGRATION OF LEUKOCYTES . . . 83 6.2.1. CD99 SUPPORTS LEUKOCYTE A D H E SIO N .............................................................. 83 6.2.2. NEITHER CD99 NOR CD99L2 ARE INVOLVED IN INTRALUMINAL CRAWLING OF LEUKO CYTES ..................................................................................................................... 85 6.2.3. CD99 AS WELL AS CD99L2 ARE INVOLVED IN ABLUMINAL C RAW LING .................... 85 6.2.4. CD99L2 IS IMPORTANT FOR THE MIGRATION THROUGH LAMININ A 5 LERS . . . 88 6.2.5. SIMULTANEOUS KNOCKOUT OF CD99 AND CD99L2 LEADS TO A TWO STEP DEFECT IN LEUKOCYTE T RA N SM IG RA TIO N .......................................................................... 89 6.3. CD99L2 GENE INACTIVATION DECREASES THE SEVERITY OF E A E ..................................... 90 6.4. FUTURE PERSPECTIVES......................................................................................................... 91 7. PUBLICATIONS 93 APPENDIX 94 A. LIST OF ACRONYMS 95 B. LIST OF TABLES 99 C. LIST OF FIGURES 100 D. BIBLIOGRAPHY 102 DANKSAGUNG 124 CURRICULUM VITAE 125
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author	Schäfer, Kerstin 1989-
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spelling	Schäfer, Kerstin 1989- Verfasser (DE-588)1174695676 aut The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall vorgelegt von Kerstin Schäfer [Münster] 2018 IV, 125 Seiten Illustrationen 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2018 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1173198237/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031441192&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Schäfer, Kerstin 1989- The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall
subject_GND	(DE-588)4113937-9
title	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall
title_auth	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall
title_exact_search	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall
title_full	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall vorgelegt von Kerstin Schäfer
title_fullStr	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall vorgelegt von Kerstin Schäfer
title_full_unstemmed	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall vorgelegt von Kerstin Schäfer
title_short	The role of CD99 and CD99L2 in leukocyte diapedesis through the blood vessel wall
title_sort	the role of cd99 and cd99l2 in leukocyte diapedesis through the blood vessel wall
topic_facet	Hochschulschrift
url	http://d-nb.info/1173198237/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031441192&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT schaferkerstin theroleofcd99andcd99l2inleukocytediapedesisthroughthebloodvesselwall

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