Apoptoseresistenz und Vermeidung von Anti-Tumor-Immunität -zwei verwandte (?) Überlebensstrategien maligner Gliomzellen [Elektronische Ressource] = Resistance to apoptosis and escape from host immune surveillance - two related (?) survival mechanisms of malignant glioma cells / vorgelegt von Jörg Hinrich Wischhusen

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Informations

• Publié par: eberhard_karls_universitat_tubingen
• Publié le: 01 janvier 2004
• Nombre de lectures: 6
• Langue: English
• Poids de l'ouvrage: 8 Mo

Extrait

Apoptoseresistenz und Vermeidung von anti-Tumor-Immunität –
zwei verwandte (?) Überlebensstrategien maligner Gliomzellen


Resistance to apoptosis and escape from host immune surveillance -
two related (?) survival mechanisms of malignant glioma cells




DISSERTATION


der Fakultät für Chemie und Pharmazie
der Eberhard-Karls-Universität Tübingen


zur Erlangung des Grades eines Doktors
der Naturwissenschaften


2004


vorgelegt von

Jörg Hinrich Wischhusen
2















Tag der mündlichen Prüfung: 14. Oktober 2004

Dekan: Professor Dr. S. Laufer
1. Berichterstatter: Professor Dr. Michael Weller
2. Berichterstatter: Professor Dr. Hans-Georg Rammensee
3. Berichterstatter: Professor Dr. Peter H. Krammer 3






















There are two rules for a good scientific presentation:
1.) Never tell everything you know...


Accordingly, the projects summarized in this thesis sought to explore and possibly overcome
human glioma cells´ resistance to apoptosis as well as their escape from host immune
surveillance. Work from other areas to which I also contributed has been omitted (see
attached reference list). 4
Proprietary Declaration

The work presented here has been performed under the guidance of Prof. Dr. Michael Weller
from January 2000 until May 2004 in the Department of Neurology at the University of
Tübingen Medical School. I hereby declare that this PhD thesis is the product of my own
work and has been written autonomously. Wherever further resources have been used, this has
been indicated. In all sections that contain work from my colleagues, I have specified the
extent of my contribution. Parts of this work have already been published and are referenced
accordingly, others have been submitted for publication.

5
Table of Contents

List of Abbreviations............................................................................................................ 9
Introductory Remark......................................................................................................... 14

I. Introduction......................................................................................................................... 15

Glioma biology and therapy.............................................................................................. 15

Prognosis and current treatment....................................................................................... 15
Perspectives...................................................................................................................... 16

Apoptosis:............................................................................................................................ 17

Physiological significance................................................................................................ 17
Mechanisms (I): the intrinsic pathway............................................................................. 18 s (II): the extrinsic pathway........................................................................... 20
Apoptosis in cancer therapy ............................................................................................. 23

Tumour immunology and immunotherapy:.................................................................... 24

The concept of immune surveillance – a historical overview.......................................... 24
Basic functions of the immune system 25
Innate immunity ............................................................................................................... 26
Adaptive immunity (I): T cells......................................................................................... 27
Antigen recognition by T cells 27
T cell development ........................................................................................................... 29
Adaptive immunity (II): antigen-presenting cells ............................................................ 30
Tumour cell recognition by the adaptive immune system ............................................... 32
Cancer immunotherapy: current strategies....................................................................... 34 clinical reponses 37
Cancer immunoediting and immune escape..................................................................... 38
Glioma-dependent immunosuppression........................................................................... 38

Experimental overview ...................................................................................................... 40

Induction of glioma cell death by rescue of mutant p53 using the experimental drug
CP-31398.......................................................................................................................... 40
Activating the extrinsic apoptotic pathway via CD40 ..................................................... 41
Glioma-induced T cell apoptosis via CD70/CD27 as a mechanism for immune escape. 41 a-dependent immunosuppression through expression of the non-classical
MHC class I molecule HLA-G......................................................................................... 42
Transforming growth factor(TGF)-β as a principal target for the immunotherapy of
malignant gliomas ............................................................................................................ 426
II. CP-31398, a novel p53-stabilizing agent, induces p53-dependent and p53-
independent glioma cell death....................................................................................... 44

Introduction ........................................................................................................................ 45
Materials and Methods ...................................................................................................... 47
Results ................................................................................................................................. 52

CP-31398 is cytotoxic to human malignant glioma cell lines.......................................... 52
Kinetic dissociation of p53-dependent and p53-independent effects of CP-31398 ......... 56
CP-31398-induced effects on malignant glioma cells: the role of the p53 status............ 63
Modulation of proapototic p53 response gene expression by CP-31398......................... 65
Modulation of Bcl-2 family protein expression by CP-31398......................................... 66
p53-independent CP-31398-induced cell death involves epiphenomenal free radical
formation, is inhibited by aurintricarboxylic acid (ATA) and may be mediated by
calcium cytotoxicity ......................................................................................................... 67
Discussion............................................................................................................................ 71

III. p53 and its family members - reporter genes may not see the difference .................. 74

Introduction ........................................................................................................................ 75
Materials and Methods ...................................................................................................... 76
Results ................................................................................................................................. 78
Discussion............................................................................................................................ 83


IV. Death receptor-mediated apoptosis in human malignant glioma cells: modulation by
the CD40/CD40L system........................................................................................................ 84

Introduction ........................................................................................................................ 85
Materials and Methods ...................................................................................................... 86
Results ................................................................................................................................. 92

CD40 expression in human glioma cell lines................................................................... 92
CD40 expression in primary human glioblastoma tissue................................................. 95
Human glioma cell lines are refractory to CD40L-mediated signaling ........................... 97
Gene transfer mediated overexpression of CD40 confers sensitivity to
CD40L-induced cell death ............................................................................................... 99
Gene transfer mediated overexpression of CD40 confers resistance to
CD95L-induced cell death ............................................................................................. 102
CD40 interacts with