Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | German |
| Veröffentlicht: |
Berlin
Mensch-und-Buch-Verl.
2001
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zugl.: Berlin, Humboldt-Univ., Diss., 2001. - Erscheint: 12. Dezember 2001 |
| Beschreibung: | 156 S. |
| ISBN: | 3898202909 |
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Datensatz im Suchindex
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| adam_text | Table of contents
Table of contents
TABLE OF CONTENTS 1
ABBREVIATIONS 6
1. ZUSAMMENFASSUNG . . 9
2. ABSTRACT 11
3. LITERATURE REVIEW 13
3.1. Chlamydial taxonomy 13
3.2. Developmental cycle of chlamydiae 15
3.3. Medical significance of chlamydiae 17
3.3.1. Chlamydia psittaci and C. pecorum 17
3.3.2. Chlamydia trachomatis 17
3.3.3. Chlamydia pneumoniae 18
3.3.3.1. Respiratory infections 18
3.3.3.2. Nonrespiratory infections 18
3.4. Atherosclerosis............................................................................................................... 19
3.4.1. Formation of atherosclerosis (atherogenesis) 19
3.4.2. Chlamydia pneumoniae, a possible etiological agent 19
3.5. Biogenesis and maturation of phagosomes 21
3.6. Fate of intracellular pathogens..................................................................................... 22
3.6.1. Pathogens residing in fusogenic vacuoles 22
3.6.2. Pathogens residing in nonfusogenic vacuoles 23
3.6.3. Pathogens escaping from vacuoles 24
3.7. Importance of iron to living cells ~™ 24
3.8. Iron acquisition systems 24
3.8.1. In eukaryotic host cells 24
Table of contents
3.8.2. In pathogens 25
3.8.2.1. Siderophore mediated iron acquisition 26
3.8.2.2. Competition for the iron loaded host proteins 26
3.8.2.2.1. Competition for Tf and lactoferrin 26
3.8.2.2.2. Competition for heme and hemoglobin 27
3.8.2.3. Iron scavenging from the labile iron pool 27
3.8.2.4. Iron acquisition by members of mammalian and bacterial NRAMP proteins... 28
3.9. Association between iron level and atherosclerosis. 28
4. MATERIALS 30
4.1. Bacterial isolates ..30
4.2. Eukaryotic host cells 30
4.3. Cell culture media 30
4.3.1. Cell growth medium (CGM) 30
4.3.2. Maintenance medium (MM) 30
4.3.3. Infection medium (IM) 31
4.4. Antibodies and fluorescent probes 31
4.5. Chemicals.... 33
4.6. Buffers and solutions .. 34
4.7. Equipment and disposable materials •• 36
5. METHODS 38
5.1. Eukaryotic cell culturing..... 38
5.2. Propagation and purification of chlamydiae.... 38
5.2.1. Bacterial growth and serial passages in cell culture 38
5.2.1.1. Chlamydia pneumoniae 38
5.2.1.2. Chlamydia trachomatis 39
5.2.2. Purification of bacteria 39
5.2.2.1. Chlamydia pneumoniae 39
Table of contents
5.2.2.2. Chlamydia trachomatis 40
S3. Titration of infectivity of purified bacteria 40
5.3.1. Infectivity titer of C. pneumoniae 40
5.3.2. Infectivity titer of C. trachomatis 41
5.4. Infection of epithelial cells for studying intracellular trafficking pattern of C.
pneumoniae..................................................................................................................... 41
5.5. Infection of epithelial cells for studying intracellular trafficking pattern of N.
gonorrhoeue 42
5.6. Immunofluorescence microscopy................................................................................ 42
5.6.1. Single staining of chlamydiae 42
5.6.1.1. Labeling using direct fluorescent method 42
5.6.1.2. Labeling using indirect fluorescent method 43
5.6.2. Double labeling of bacteria and endocytic markers 43
5.7. Texas Red dextran labeling............. ...... ........ .......... ......... .............. 44
5.8. Estimation of the pH of C. pneumoniae containing vacuoles......................... ...... 44
5.8.1. Acridine Orange and LysoTracker staining 44
5.8.2. DAMP staining 45
5.9. Treatment with the acidification inhibitor bafilomycin Al 46
5.10. Treatment with chloramphenicol 46
5.11. Withholding of host intracellular iron and C. pneumoniae infection assays 47
5.11.1. Infection of cells with C. pneumoniae under iron depletion conditions 47
5.11.2. Effect of iron depletion on morphology of C. pneumoniae inclusion 47
5.11.3. Effect of iron depletion on C. pneumoniae infectivity 48
5.12. Withholding of host intracellular iron and C. trachomatis infection assays 48
5.12.1. Infection of cells with C. trachomatis under iron depletion conditions 48
5.12.2. Effect of iron depletion on morphology of C. trachomatis inclusion 49
5.12.3. Effect of iron depletion on C. trachomatis infectivity 49
5.13. Influence of HTf on C. pneumoniae growth 49
Table of contents
5.14. Host cell viability 50
5.15. Transmission electron microscopy............................................................................. 51
5.16. One dimensional gel electrophoresis and immunoblotting....................................... 51
5.16.1. SDS PAGE 51
5.16.2. Western blot 52
5.16.2.1. Protein transfer 52
5.16.2.2. Immunoblotting 52
5.17. Statistical analysis 53
6. PART ONE:
CHARACTERIZATION AND INTRACELLULAR TRAFFICKING PATTERN OF
VACUOLES CONTAINING C. PNEUMONIAE IN HUMAN EPITHELIAL CELLS. 55
6.1. Introduction and goals................................................................................................— 56
6.2. Results ..................................................................................... ...,.........„.....••..¦•••••• 58
6.2.1. Intravacuolar nature of C. pneumoniae inclusion 58
6.2.1.1. Qualitative assessment of the pH of the inclusion 58
6.2.1.2. Effect of the acidification inhibitor bafilomycin Al on chlamydial growth 62
6.2.2. Interaction of the host endocytic organelles with C. pneumoniae inclusion 66
6.2.3. Early endosomes are intimately associated with C. pneumoniae vacuole 66
6.2.4. Chlamydia pneumoniae inclusion does not interact with late endosomes 68
6.2.5. Chlamydia pneumoniae conlainmg inclusion avoids fusion with lysosomes 70
6.2.6. The effect of chlamydial protein synthesis inhibition on the interaction with
lysosomes 74
63. Discussion . . . .... .... . .. ........79
7. PART TWO:
LOW IRON AVAILABILITY MODULATES THE COURSE OF C. PNEUMONIAE
INFECTION 85
7.1. Introduction and goals 86
Table of contents
7.2. Results 88
7.2.1. Effects of iron chelation on C. trachomatis L2 growth 88
7.2.1.1. Iron limitation affects the size of the inclusion 88
7.2.1.2. Iron limitation delays the homotypic inclusion fusion 88
7.2.1.3. Effect of iron limitation on C. trachomatis infectivity 91
7.2.2. Effects of iron chelation on C. pneumoniae growth 91
7.2.2.1. Iron limitation affects the size of the inclusion 91
7.2.2.2. Effect of iron limitation on C. pneumoniae infectivity 95
7.2.3. Ultrastructural analysis of C. pneumoniae infected cells deprived of iron 95
7.2.4. Exogenous iron saturated Tf restores the growth of iron deprived C. pneumoniae 98
7.2.5. Interaction of iron starved C. pneumoniae vacuole with endocytic pathway of host
cells 101
7.2.6. Chlamydia pneumoniae infection does not affect amounts of cellular TfR 101
7.2.7. Capacity of iron limitation to develop chlamydial persistence 103
73. Discussion HO
8. REFERENCES 117
CURRICULUM VITAE 144
PUBLICATIONS 145
ACKNOWLEDGEMENT 147
SELBSTANDIGKEITSERKLARUNG 149
5
|
| any_adam_object | 1 |
| author | Al-Younes, Hesham 1967- |
| author_GND | (DE-588)123404312 |
| author_facet | Al-Younes, Hesham 1967- |
| author_role | aut |
| author_sort | Al-Younes, Hesham 1967- |
| author_variant | h a y hay |
| building | Verbundindex |
| bvnumber | BV014064032 |
| classification_rvk | WF 6000 |
| ctrlnum | (OCoLC)633335141 (DE-599)BVBBV014064032 |
| discipline | Biologie |
| format | Book |
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| indexdate | 2024-07-09T18:57:00Z |
| institution | BVB |
| isbn | 3898202909 |
| language | German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009632549 |
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| spelling | Al-Younes, Hesham 1967- Verfasser (DE-588)123404312 aut Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development von Hesham Al-Younes Berlin Mensch-und-Buch-Verl. 2001 156 S. txt rdacontent n rdamedia nc rdacarrier Zugl.: Berlin, Humboldt-Univ., Diss., 2001. - Erscheint: 12. Dezember 2001 Eisenmangel (DE-588)4151569-9 gnd rswk-swf Chlamydia pneumoniae (DE-588)4367505-0 gnd rswk-swf Immunfluoreszenz (DE-588)4161372-7 gnd rswk-swf Wasserstoffionenkonzentration (DE-588)4189273-2 gnd rswk-swf Vakuole (DE-588)4187316-6 gnd rswk-swf Epithelzelle (DE-588)4259648-8 gnd rswk-swf Eisen (DE-588)4014002-7 gnd rswk-swf Wachstum (DE-588)4064115-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Chlamydia pneumoniae (DE-588)4367505-0 s Eisenmangel (DE-588)4151569-9 s Wachstum (DE-588)4064115-6 s DE-604 Vakuole (DE-588)4187316-6 s Immunfluoreszenz (DE-588)4161372-7 s Epithelzelle (DE-588)4259648-8 s Wasserstoffionenkonzentration (DE-588)4189273-2 s Eisen (DE-588)4014002-7 s HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009632549&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Al-Younes, Hesham 1967- Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development Eisenmangel (DE-588)4151569-9 gnd Chlamydia pneumoniae (DE-588)4367505-0 gnd Immunfluoreszenz (DE-588)4161372-7 gnd Wasserstoffionenkonzentration (DE-588)4189273-2 gnd Vakuole (DE-588)4187316-6 gnd Epithelzelle (DE-588)4259648-8 gnd Eisen (DE-588)4014002-7 gnd Wachstum (DE-588)4064115-6 gnd |
| subject_GND | (DE-588)4151569-9 (DE-588)4367505-0 (DE-588)4161372-7 (DE-588)4189273-2 (DE-588)4187316-6 (DE-588)4259648-8 (DE-588)4014002-7 (DE-588)4064115-6 (DE-588)4113937-9 |
| title | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development |
| title_auth | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development |
| title_exact_search | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development |
| title_full | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development von Hesham Al-Younes |
| title_fullStr | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development von Hesham Al-Younes |
| title_full_unstemmed | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development von Hesham Al-Younes |
| title_short | Characterization of vacuoles containing Chlamydia pneumoniae (synonym Chlamydophila pneumoniae) in human epithelial cells and the role of iron in the pathogen development |
| title_sort | characterization of vacuoles containing chlamydia pneumoniae synonym chlamydophila pneumoniae in human epithelial cells and the role of iron in the pathogen development |
| topic | Eisenmangel (DE-588)4151569-9 gnd Chlamydia pneumoniae (DE-588)4367505-0 gnd Immunfluoreszenz (DE-588)4161372-7 gnd Wasserstoffionenkonzentration (DE-588)4189273-2 gnd Vakuole (DE-588)4187316-6 gnd Epithelzelle (DE-588)4259648-8 gnd Eisen (DE-588)4014002-7 gnd Wachstum (DE-588)4064115-6 gnd |
| topic_facet | Eisenmangel Chlamydia pneumoniae Immunfluoreszenz Wasserstoffionenkonzentration Vakuole Epithelzelle Eisen Wachstum Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009632549&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT alyouneshesham characterizationofvacuolescontainingchlamydiapneumoniaesynonymchlamydophilapneumoniaeinhumanepithelialcellsandtheroleofironinthepathogendevelopment |