Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 306 Towards Immunotherapy of Midgut Carcinoid Tumors SOFIA VIKMAN ACTA UNIVERSITATIS UPSALIENSIS ISSN 1651-6206 UPPSALA ISBN 978-91-554-7076-0 2008 urn:nbn:se:uu:diva-8421 ! "#$% & ' & & (' ' ) & * +, -' . / ', 0 1, !, - . 2 ' & * 3 - , 4 , 56, %6 , , 2178 "9!:"$:%%;:996:, 3 & ' )</(:8/-+ .' , -' & ' = & # ' && ' . ' , ' ' ' > & ' : , -' '> &' ' ' ' , - & && - ' ' ', ? & : & ' , 3' 4 )3<4+ ' ' $ )-(:$+ $ )0*4-:$+ ' & )3@:+ )24:+ ' & , 2 & . & & 0*4-:$ ' , -' 0*4-$A$% && 3: ' & > ' )/+ & , ? & B4:4C$: & > 3!D - & , ? & & 3!D - > ' & '' , ( ' &E & - )-+ .' E ' & '' , * &E & - ' . ' & 5: -, * : > 3!D - & & ' ' & 3 - &, . ' & -'$ && ' , ' 0*4-:$ - B4:4C$: 2 8F - ! " # $ % &'()*+) G 1 & 0 ! 2118 $6%$:66 2178 "9!:"$:%%;:996: # ### :!;$ )' #HH ,,H I J # ### :!;$+ g|ÄÄ Å|Ç ytÅ|Ä} List of papers This thesis is based on the following papers, which will be referred to in the text by their roman numerals: I Vikman S., Essand M., Cunningham JL., de la Torre M., Öberg K., Tötterman TH. and Giandomenico V. Gene Expression in Mid- gut Carcinoid Tumors: Potential Targets for Immunotherapy. Acta Oncologica 2005; 44(1):32-40. II Essand M., Vikman S., Grawé J., Gedda L., Hellberg C., Öberg K., Tötterman TH. and Giandomenico V. Identification and Characterization of a Novel Splicing Variant of Vesicular Mono- amine Transporter 1. Journal of Molecular Endocrinology 2005; 35(3):489-501. III Vikman S., Giandomenico V., Sommaggio R., Öberg K., Essand M. and Tötterman TH. CD8+ T cells against Multiple Tumor- associated Antigens in Peripheral Blood of Midgut Carcinoid Pa- tients. Cancer Immunology, Immunotherapy 2008; 57(3): 399-409. IV Vikman S., Sommaggio R., de la Torre M., Öberg K., Essand M., Giandomenico V., Loskog A. and Tötterman TH. Midgut Carci- noid Patients Display Increased Numbers of Regulatory T cells in Peripheral Blood with Infiltration into Tumor Tissue. Manuscript. Reprints were made with permission from the publishers I Copyright © 2005 Taylor & Francis Group II Copyright © 2005 Society for Endocrinology III Copyright © 2008 Springer Science & Business Media Contents Introduction...................................................................................................13 The dispersed neuroendocrine system......................................................13 General overview.................................................................................13 Neuroendocrine cellular markers.........................................................13 Neuroendocrine cells of the gastrointestinal tract................................14 Classical midgut carcinoid tumors.......................................................16 The immune system .................................................................................18 General overview.................................................................................18 Antigen presentation............................................................................19 Dendritic cells......................................................................................21 T cell activation and tolerance.............................................................23 Tumor immunology..................................................................................26 History of tumor immunology.............................................................26 Immune recognition of tumors ............................................................27 Immune escape mechanisms used by tumors ......................................28 Tumor antigens....................................................................................30 Tumor antigen discovery .....................................................................31 T cell immunotherapy of tumors..............................................................32 General overview.................................................................................32 Modified dendritic cell vaccines..........................................................32 Adoptive transfer of in vitro activated T cells .....................................32 Genetically engineered T cells.............................................................33 Present investigation .....................................................................................34 General aim ..............................................................................................34 Specific aims ............................................................................................34 Materials and methods .............................................................................34 Results and discussion ..................................................................................35 Paper I: Gene Expression in Midgut Carcinoid Tumors: Potential Targets for Immunotherapy......................................................................35 Paper II: Identification and Characterization of a Novel Splicing Variant of Vesicular Monoamine Transporter 1 ......................................36 Paper III: CD8+ T cells against Multiple Tumor-associated Antigens in Peripheral Blood of Midgut Carcinoid Patients ...................................38 Paper IV: Midgut Carcinoid Patients Display Increased Numbers of Regulatory T cells in Peripheral Blood with Infiltration into Tumor Tissue............................................................................................40 Conclusions...................................................................................................42 Acknowledgements.......................................................................................43 References.....................................................................................................46 Abbreviations aa Amino acid ADCC Antibody-dependent, cell-mediated cytotoxicity AINR Activation-induced, non-responsiveness APC Antigen presenting cell APUD Amine precursor uptake and decarboxylation CCL Chemokine (C-C) motif ligand CCR Chemokine (C-C) motif receptor CD Cluster of differentiation cDNA Complementary DNA CD40L CD40 ligand CDX-2 Caudal type homeobox transcription factor 2 CEA Carcinoembryonic antigen CFSE Carboxyfluorescein diacetate succinimidyl ester CGA Chromogranin A CML Chronic myeloid leukaemia CNS Central nervous system CpG Cytosine-phosphate-guanine CT Cancer-testis CT Computed tomography CTL Cytotoxic T lymphocytes CTLA-4 Cytotoxic T lymphocyte-associated antigen 4 DC Dendritic cell DNA Deoxyribonucleic acid dsRNA Double-stranded ribonucleic acid EBV Epstein-Barr virus EC Enterochromaffin EGFP Enhanced green fluorescent protein ER Endoplasmic reticulum ERAP1 Endoplasmic reticulum aminopeptidase 1 FLIP FLICE-inhibitory protein Foxp3 Forkhead box p3 GAPDH Glyceraldehyde-3-phosphate dehydrogenase GI Gastrointestinal GM-CSF Granulocyte macrophage colony stimulating factor HER-2 Human epidermal growth factor receptor 2 5-HIAA 5-Hydroxy indol acetic acid HLA Human leukocyte antigen HPV Human Papilloma virus 5-HT 5-Hydroxytryptamine IA-2 Islet autoantigen-2 IDO Indoleamine-2,3-dioxygenase IFN Interferon IgG Immunoglobulin G IL Interleukin IPEX Immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome LDCV Large dense core vesicles LFA Leukocyte function-associated antigen LPS Lipopolysaccharide M-CSF Macrophage colony stimulating factor mDC Myeloid dendritic cell MHC Major histocompatibility complex mRNA Messenger ribonucleic acid MRT Magnetic resonance tomography NCI National Cancer Institute NFAT Nuclear factor of activated T cells NFB Nuclear factor kappa B NK Natural killer PAMP Pathogen-associated molecular pattern pDC Plasmacytoid dendritic cell PET Positron emission tomography PGE2 Prostaglandin E2 PGP 9.5 Protein gene product 9.5 PI-9 Protein inhibitor 9 PRR Pattern recognition receptors RT-PCR Reverse transcription polymerase chain reaction SEREX Serological identification of antigens by recombinant expression cloning SLMV Synaptic-like microvesicles SNSD Self/Non-self discrimination TAP Transporter associated with antigen processing TCM Central memory T cells TCR T cell receptor TEM Effector memory T cells TGF Tumor growth factor TH T helper TH1 T helper type 1 TH2 T helper type 2 TH3 T helper type 3 TILs Tumor infiltrating lymphocytes TLR Toll-like receptor TNF Tumor necrosis factor TPH Tryptophan hydroxylase TR1 Regulatory type 1 cells Tregs Regulatory T cells VAT Vesicular amine transporter VEGF Vascular endothelial growth factor VMAT Vesicular monoamine transporter WB Western blot WHO World Health Organisation Introduction The dispersed neuroendocrine system General overview Neuroendocrine cells are distributed throughout most tissues in the body and they can be organized as macroscopic organs, such as the adenohypophysis and the adrenal medulla, or dispersed among other cell types as single or small groups of cells, such as the islets of Langerhans. The cells are special- ized in