Tolerance and immunity to human tumor-associated antigens [Elektronische Ressource] / Carina Lotz

Tolerance and Immunity to Human Tumor-Associated Antigens Dissertation zur Erlangung des Grades “Doktor der Naturwissenschaften” am Fachbereich Biologie der Johannes Gutenberg-Universität in Mainz Carina Lotz geboren in Diez/Lahn Mainz, 2003 Dekan: 1. Berichterstatter: 2. Berichterstatter: Tag der mündlichen Prüfung: 16.12.2003 To my parents Table of Contents I Table of Contents Abbreviations IV 1. Introduction 1 1.1 Tumor-associated antigens: The key to cytotoxic T lymphocyte 1 (CTL)-based immunotherapy of malignant disease 1.2 Self tolerance to tumor-associated antigens 5 1.3 Strategies to circumvent self tolerance 8 1.4 Specific aims 10 2. Materials and Methods 11 2.1 Mice 11 2.2 Synthetic peptides 2.3 Antibodies 12 2.4 Viruses, cells and cell lines 13 2.5 Flow cytometric analysis 15 2.6 Western blot analysis 16 2.7 Immunohistochemistry 17 2.8 Peptide-binding to HLA-A*0201 (A2.1) 18 2.9 Peptide priming of A2.1 transgenic mice, propagation of CTL 18 and depletion of T regulatory cells 2.10 Inhibiting 20S proteasomal antigen production 19 2.11 Extracting natural peptides by MHC-immunoprecipitation 19 and RP-HPLC separation 51 2.12 Cr-release and ELISPOT assay 20 2.13 Inducing A2.1-restricted and peptide-specific human CTL 22 3.