https://edoc.ub.uni-muenchen.de/5277/1/adhikary_dinesh.pdf
Investigation of the T helper cell response against
Epstein-Barr virus
A thesis submitted for the degree of Doctor of Medicine
at the Faculty of Medicine
Ludwig-Maximilians-University Munich
Dinesh Adhikary
2006
Completed at the GSF Research Center for Environment and Health
Institute for Clinical Molecular Biology and Tumour Genetics, Munich.
Aus dem GSF Forschungszentrum für Umwelt und Gesundheit,
Institut für Klinische Molekularbiologie und Tumorgenetik
Leiter des Instituts: Prof. Dr. med. Georg W. Bornkamm
Untersuchung der T-Helferzellantwort gegen
das Epstein-Barr Virus
Dissertation
zum Erwerb des Doktorgrades der Medizin
an der Medizinischen Fakultät der
Ludwig-Maximilians-Universität zu München
vorgelegt von
Dinesh Adhikary
aus
Saping village, Nepal
2006
Mit Genehmigung der medizinischen Fakultät
der Universität München
1. Berichterstatter: Prof. Dr. med. Georg W. Bornkamm
2. Mitberichterstatter: Prof. Dr. H. G. Klobeck
Prof. Dr. U. Koszinowski
Prof. Dr. Th. Brocker
Mitbetreuung durch
die promovierten Mitarbeiter: Dr. rer. nat. Josef Mautner, Dr. med. Uta Behrends
Dekan: Prof. Dr. med. D. Reinhardt
Tag der mündlichen Prüfung: 11.05.2006
Table of Contents
Table of Contents
A. Abstracts
A.1 Zusammenfassung 1
A.2 Summary 3
B. Abbreviations 5
1. Introduction 8
1.1 The Epstein-Barr virus 8
1.1.1 Classification and structure 8
1.1.2 Epidemiology and clinical manifestations 9
1.1.3 Latent and lytic infection 11
1.1.4 EBV infection in the natural host 12
1.1.5 EBV infection in vitro 14
1.2 Immune responses to EBV 15
1.2.1 Humoral immune response 15
1.2.2 The CD8+ T cell response to EBV 16
1.2.3 The CD4+ T cell response to EBV 18
1.3 EBV in the immunocompromised 21
1.4 Immunological approaches to control EBV infection 22
1.4.1 T cell therapy of EBV-associated lymphomas 23
1.5 Aim of the work 24
2. Materials and Methods 25
2.1 Laboratory equipments 25
2.2 Cell biology materials and methods 26
2.2.1 Eukaryotic cell culture materials 26
2.2.2 General conditions for eukaryotic cell culture 27
2.2.3 Separation of peripheral blood mononuclear cells from whole blood 28
2.2.4 Preparation of pooled human sera for T cell media 29
2.2.5 Establishment and culture of lymphoblastoid cell lines 29
2.2.6 Generation and maintenance of T cell lines and clones 30
2.2.7 Preparation of dendritic cells from PBMCs 31
2.2.8 Methods for T cell phenotype and function analysis 32
2.2.8.1 Flow cytometric analysis of T cells 32
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