Composition and efficacy of cytotoxic T-cell responses determine virus elimination and immunopathology after virus infections [Elektronische Ressource] / vorgelegt von Birthe Jessen

Agehl

Sujets

• Biologie

Informations

• Publié par: albert-ludwigs-universitat_freiburg
• Publié le: 01 janvier 2010
• Nombre de lectures: 14
• Langue: Deutsch
• Poids de l'ouvrage: 1 Mo

Extrait

ALBERT-LUDWIGS-UNIVERSITÄT FREIBURG IM BREISGAU


Composition and efficacy of cytotoxic
T cell responses determine virus elimination
and immunopathology after virus infections





INAUGURAL-DISSERTATION
zur Erlangung der Doktorwürde der Fakultät für
Biologie und der Fakultät für Medizin
der Albert-Ludwigs-Universität
Freiburg im Breisgau


vorgelegt von
Birthe Jessen
aus Bad Neustadt/Saale

September 2010


Dekan der Biologischen Fakultät: Prof. Dr. rer. nat. Ad Aertsen

Dekan der Medizinischen Fakultät: Prof. Dr. med. Christoph Peters

Betreuer der Arbeit/Doktorvater: Prof. Dr. Hanspeter Pircher

Betreuer der Arbeit: Prof. Dr. Stephan Ehl

Koreferent: Prof. Dr. Peter Stäheli

Promotionsvorsitzender: Prof. Dr. Samuel Rossel



Tag der Verkündigung des Prüfungsergebnisses: 30.11.2010









Diese Arbeit wurde am Centrum für Chronische Immundefizienz (CCI) des
Universitätsklinikums Freiburg - Albert-Ludwigs-Universität Freiburg - erstellt.




























‘Nothing shocks me. I'm a scientist.’

- Harrison Ford as Indiana Jones Contents 4
Contents............................................................................................................................... 4
Abstract ............................................................................................................................... 7
Abbreviations ...................................................................................................................... 8

1 Introduction ..................................................................................................... 10

1.1 Immune system...................................................................................................10
1.1.1 Innate immune system ................................................................................................ 10
1.1.2 Adaptive immune response......................................................................................... 11
1.1.3 Antiviral immune responses ........................................................................................ 12
1.2 T cell-mediated immunopathology following RSV infection............................13
1.3 Control of immune homeostasis by T cells ......................................................14
1.4 Cell death induced by cytotoxic lymphocytes ..................................................15
1.4.1 Ligation of death receptors.......................................................................................... 15
1.4.2 Exocytosis of lytic granules ......................................................................................... 16
1.5 Hemophagocytic Lymphohistiocytosis.............................................................19
1.5.1 Genetic defects affecting lymphocyte cytotoxicity....................................................... 19
1.5.1.1 Familiar hemophagocytic lymphohistiocytosis (FHL).............................................. 21
1.5.1.2 Chèdiak-Higashi syndrome..................................................................................... 22
1.5.1.3 Griscelli syndome type II......................................................................................... 23
1.5.1.4 Hermansky-Pudlak syndrome type II ...................................................................... 24
1.5.2 Diagnostic criteria........................................................................................................ 25
1.5.3 Treatment .................................................................................................................... 26
1.5.4 Open questions in disease pathogenesis ................................................................... 26
1.6 Aims of the study................................................................................................29

2 Materials and Methods.................................................................................... 30

2.1 Mice, Viruses and Materials ...............................................................................30
2.1.1 Mice ............................................................................................................................. 30
2.1.2 Viruses......................................................................................................................... 30
2.1.3 Cells............................................................................................................................. 31
2.1.4 Narcotics...................................................................................................................... 31
2.1.5 Cell culture media........................................................................................................ 31
2.1.6 Synthetic peptides ....................................................................................................... 32
2.1.7 Antibodies.................................................................................................................... 32
2.1.8 Primer .......................................................................................................................... 33
2.1.9 Kits............................................................................................................................... 34
2.1.10 Enzymes...................................................................................................................... 35 Contents 5
2.1.11 Chemicals, buffers and solutions ................................................................................ 35
2.1.12 Plastic materials .......................................................................................................... 37
2.1.13 Instruments.................................................................................................................. 38
2.2 Methods...............................................................................................................39
2.2.1 Viruses......................................................................................................................... 39
2.2.2 Hybridoma ................................................................................................................... 39
2.2.3 Mice ............................................................................................................................. 39
2.2.5 Treatment of mice........................................................................................................ 43
2.2.6 Preparation of mice ..................................................................................................... 44
2.2.7 In vitro activation of T cells .......................................................................................... 44
2.2.8 Determination of virus titers......................................................................................... 45
2.2.9 Flow cytometry ............................................................................................................ 46
2.2.10 Magnetic Activated Cell Separation ............................................................................ 47
2.2.11 Blood count.................................................................................................................. 47
2.2.12 Proliferation assay....................................................................................................... 47
2.2.13 Cytotoxicity Assay ....................................................................................................... 47
2.2.14 Determination of cytokine levels.................................................................................. 48
2.2.15 Analysis of liver enzymes, triglycerides and ferritin serum levels ............................... 49
2.2.16 Histology...................................................................................................................... 49
2.2.17 Statistical analysis ....................................................................................................... 49

3 Results ............................................................................................................. 50

3.1 Strain-specific disease susceptibility after RSV infection in the mouse is
determined by MHC dependent CTL responsiveness ......................................50
3.1.1 The MHC haplotype is an important determinant of disease susceptibility following
RSV infection. .............................................................................................................. 50
3.1.2 RSV induced disease is not determined by peak virus titers or virus elimination
kinetics. ........................................................................................................................ 51
3.1.3 The pulmonary CTL response is of similar magnitude in MHC congenic mice........... 52
+3.1.4 Neither re