Enhancing anti-tumor immunity to MHC class I-deficient tumors [Elektronische Ressource] : role of regulatory T cells and type I IFN / presented by Ioanna Evdokia Galani

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Sujets

• Biologie

Informations

• Publié par: ruprecht-karls-universitat_heidelberg
• Publié le: 01 janvier 2008
• Nombre de lectures: 6
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

Enhancing anti-tumor immunity to MHC class I-deficient
tumors: role of regulatory T cells and type I IFN





Dissertation
submitted to the
Combined Faculties for the Natural Sciences and for Mathematics
of the Ruperto-Carola University Of Heidelberg, Germany
for the degree of
Doctor of Natural Sciences










presented by
Ioanna Evdokia Galani
born in Acharnes, Greece

Dissertation
submitted to the
Combined Faculties for the Natural Sciences and for Mathematics
of the Ruperto-Carola University Of Heidelberg, Germany
for the degree of
Doctor of Natural Sciences















presented by
Ioanna Evdokia Galani
born in Acharnes, Greece


Oral examination: 20.06.2008



Enhancing anti-tumor immunity to MHC class I-deficient
tumors: role of regulatory T cells and type I IFN





















1. Referee: Prof. Dr. G. Hämmerling
2. Referee: PD Dr. A. Cerwenka


The work described in this thesis was performed from September 2004 to April 2008 at the
German Cancer Research Center, Germany, in the laboratory of PD Dr. Adelheid Cerwenka.


Parts of this thesis have been published in:

Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. Treg suppress leukocyte
accumulation and macrophage activation in lymphoma. Manuscript in preparation.

Nausch N., I.E. Galani and A. Cerwenka. Myeloid-derived ‚suppressor’ cells express RAE-1 and
activate NK cells. Manuscript in revision.

Wendel M., I.E. Galani, E. Suri-Payer and A. Cerwenka. NK cell accumulation in tumors is
dependent on IFN- γ and CXCR3 ligands. Manuscript submitted.

Kiessling M., C.D. Klemke, M. Kami ński, I.E. Galani, P.H. Krammer and K. Gülow. Survival of
primary T cell lymphomas is dependent on regulation of Ferritin Heavy Chain by constitutively
activated NF- κB. Manuscript submitted.


Conference presentations:

Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. IFN- γ dependent rejection
thof the NK cell dependent RMA-S tumor in the absence of regulatory T cells. 16 European
Congress of Immunology, September 2006, Paris, France.

Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. Regulatory T cells
suppress IFN- γ dependent rejection of the NK cell sensitive RMA-S tumor. Natural Killer Cell
Symposium 2006 Heidelberg, December 2006, Heidelberg, Germany.
Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. Rejection of the NK cell
sensitive tumor RMA-S via IFN- γ in the absence of regulatory T cells. Keystone Symposia
meeting ‘Mechanisms Linking Inflammation and Cancer’, February 2007, Santa Fe, New
Mexico, USA.

Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. Treg suppress leukocyte
thaccumulation and macrophage activation in RMA-S lymphoma. 10 Meeting of the Society for
Natural Immunity, April 2007, Cambridge, UK.

Galani I.E., M. Wendel, C. Schellack, E. Suri-Payer and A. Cerwenka. Regulatory T cells
th suppress IFN- γ dependent leukocyte accumulation and macrophage activation in lymphoma. 37
Annual Meeting of the German Society for Immunology (DGfI), September 2007, Heidelberg,
Germany.


















CONTENTS

1 ZUSAMMENFASSUNG .......................................................................................... 1
2 SUMMARY................................................................................................................ 3
3 INTRODUCTION..................................................................................................... 5
3.1 THE IMMUNE SYSTEM....................................................................................................5
3.1.1 The immune response to tumors ............................................................................5
3.1.2 The innate immune system.....................................................................................7
3.1.2.1 Natural Killer cells...........................................................................................7
3.1.2.1.1 NK cells and tumor......................................................................................11
3.1.2.1.2 NK cells as regulatory cells.........................................................................13
3.1.2.2 Macrophages..................................................................................................14
3.1.2.2.1 Tumor associated macrophages...................................................................15
3.1.2.2.2 Myeloid-Derived Suppressor Cells.............................................................16
3.1.3 The adaptive immune system...............................................................................17
3.1.3.1 Mechanisms of T cell tolerance.....................................................................18
3.1.3.2 Regulatory T cells..........................................................................................20
3.1.3.2.1 Phenotypic characterization of Treg............................................................20
3.1.3.2.2 Treg and autoimmunity .........................................22
3.1.3.2.3 Treg and tumor immunity.........................................................22
3.1.3.2.4 Treg and innate immunity.........................23
3.1.3.2.5 Manipulation of the suppressive function of Treg.......................................24
3.2 TYPE I IFN.....................................................................................................................25
3.2.1 Regulatory effects of type I IFN...........................................................................26
3.2.2 Type I IFN and tumor...........................................................................................27
4 AIM OF THE STUDY.....................................................................................................29
5 MATERIAL AND METHODS .....................................................................................30
5.1 MATERIALS..................................................................................................................30
5.1.1 Laboratory equipment..........................................................................................30 5.1.2 Cell culture products ............................................................................................31
5.1.3 Cell culture media................................................................................................32
5.1.4 Solutions...............................................................................................................33
5.1.5 Chemicals.............................................................................................................34
5.1.6 Antibodies............................................................................................................35
5.1.6.1 Antibodies for FACS analysis .......................................................................35
5.1.6.2 Antibodies for in vitro activation...................................................................35
5.1.6.3 In vivo administered antibodies .....................................................................35
5.1.7 Cell lines...............................................................................................................38
5.1.8 Magnetic Cell Sorting (MACS) beads and columns ............................................38
5.1.9 Kits.......................................................................................................................39
5.2 MICE .............................................................................................................................40
5.3 METHODS .....................................................................................................................40
5.3.1 Cell culture methods.............................................................................................40
5.3.1.1 Determination of cell number........................................................................40
5.3.1.2 Freezing and thawing of cells ........................................................................40
5.3.1.3 Splitting of adherent cells ..............................................................................41
5.3.1.4 Splitting of suspension cells ..........................................................................41
5.3.2 Mouse tumor model .............................................................................................41
5.3.2.1 Isolation of organs and preparation of single cell suspensions......................42
5.3.2.1.1 Blood...........................................................................................................42
5.3.2.1.2 Spleen and LN .............................................................................................42
5.3.2.1.3 Tumor inf