Establishment of in vivo bioluminescence imaging models for tumor immunology [Elektronische Ressource] / presented by Tewfik Miloud

Miloud Zaater

Sujets

• Gesundheit

Informations

• Publié par: ruprecht-karls-universitat_heidelberg
• Publié le: 01 janvier 2008
• Nombre de lectures: 74
• Langue: English
• Poids de l'ouvrage: 16 Mo

Extrait

Establishment of in vivo bioluminescence imaging
models for tumor immunology








InauguralDissertation
Tewfik Miloud
























































DISSERTATION


Submittedtothe
FakultätfürBiowissenschaftenof
RuprechtKarlUniversität
Heidelberg

andtothe

UFRsciencesdelavieof
UniversitédeBourgogne
Dijon



Presentedby
Tewfik Miloud
Bornin:LeCreusot,France
Oralexamination:23rdofNovember2007







Establishment of in vivo bioluminescence imaging
models for tumor immunology












Supervisors:
Prof.Dr.GünterJ.Hämmerling
Prof.JohannaChluba

Reviewers:
Dr.Protzer
Dr.Aprahamian
Table of contents

List of figures: .................................................................................................... vii

List of tables: ..................................................................................................... viii

Publications: ........................................................................................................ ix

Summary .............................................................................................................. x
A. Introduction .................................................................................. 1
1. Imaging.........................................................................................................2
1.1 Tumor imaging in mouse model of cancer...................................................................2
1.1.1 MRI................................................................................................................................................2
1.1.2 CT...................................................................................................................................................3
1.1.3 US...................................................................................................................................................3
1.1.4 PET and SPECT............................................................................................................................3
1.2 Optical imaging...............................................................................................................4
1.2.1 Bioluminescence imaging..............................................................................................................5
1.2.1.1 Bioluminescence.....................................................................................................................5
1.2.1.2 Principle of Bioluminescence imaging..................................................................................5
1.2.2 Luciferases.....................................................................................................................................6
1.2.2.1 Firefly luciferase.....................................................................................................................6
1.2.2.2 Renilla luciferase....................................................................................................................7
1.2.2.3 Bacterial luciferase (lux operon)............................................................................................7
1.2.3 Light transmission through mammalian tissue..........................................................................7
1.2.4 Use of BLI in monitoring biological processes............................................................................8

2. Animal models for tumor immunology.....................................................9
2.1 Transplantable tumor models........................................................................................9
2.2 Autochthonous tumor models......................................................................................10
2.2.1 SV40-driven transgenic models.................................................................................................11
2.2.2 Inducible gene expression systems.............................................................................................11
2.2.2.1 Cre/LoxP system...................................................................................................................12
2.2.2.2 Conditional liver tumor model..............................................................................................15



i Table of contents
3. Bacteria and anticancer therapies...........................................................15
3.1 Bacteria and tumor colonization.................................................................................15
3.2 Why do bacteria colonize tumors?..............................................................................16
3.3 Bacteria as a vaccine vehicle........................................................................................17

4. Aims of the study.......................................................................................18
B. Material and methods ................................................................ 19
1. Material......................................................................................................20
1.1 Chemicals.......................................................................................................................20
1.2 Basic equipment............................................................................................................20
1.3 Kits.................................................................................................................................20
1.4 Technical devices...........................................................................................................21
1.5 Buffers and solutions....................................................................................................22
1.6 Media..............................................................................................................................25
1.6.1 Media for bacterial culture........................................................................................................25
1.6.2 Media for cell culture..................................................................................................................25
1.7 Bacterial strains............................................................................................................26
1.8 Mammalian cell lines....................................................................................................26
1.9 Mouse lines....................................................................................................................26
1.10 Antibodies......................................................................................................................27
1.11 Plasmids.........................................................................................................................27
1.12 PCR primers..................................................................................................................28

2. Methods......................................................................................................29
2.1 Molecular biology.........................................................................................................29
2.1.1 Bacterial culture..........................................................................................................................29
2.1.2 Preparation of CaCl competent (heat competent) E. coli bacteria........................................292
2.1.3 Bacteria storage...........................................................................................................................29
2.1.4 Transformation of heat (CaCl ) competent E. coli bacteria....................................................292
2.1.5 DNA minipreparation (alkaline lysis).......................................................................................30
2.1.6 DNA midipreparation (alkaline lysis).......................................................................................30
2.1.7 Restriction digestion...................................................................................................................31
2.1.8 Dephosphorylation of DNA ends (e.d. in vectors for ligation).................................................31
ii Table of contents
2.1.9 Electrophoretic separation of DNA fragments in agarose gels...............................................31
2.1.10 Isolation of DNA fragments from agarose gels with Qiaquick Gel extraction kit...............32
2.1.11 Ligation of DNA fragments and vectors..................................................................................32
2.1.12 PCR (Polymerase Chain Reaction)