by Chan Ka Lee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 94-112). / Abstracts in English and Chinese. / Acknowledgements --- p.i / abstract(english/chinese) --- p.iii / Contents --- p.vii / List of Tables --- p.xi / List of Figures --- p.xii / Chapter I --- Introduction --- p.1 / Chapter I.1 --- Gastric Lymphoma --- p.1 / Chapter I.1.1 --- Background --- p.1 / Chapter I.1.2 --- Mucosa-Associated Lymphoid Tissue --- p.2 / Chapter I.1.3 --- Classification of Primary Gastric Lymphomas --- p.3 / Chapter I.1.3.1 --- Mucosa-Associated Lymphooid Tissue Type Lymphomas --- p.3 / Chapter I.1.3.2 --- High Grade Primary Gastric Lymphomas --- p.5 / Chapter I.1.3.3 --- Other Gastric Lymphomas --- p.7 / Chapter I.2 --- Helicobater Pylori --- p.8 / Chapter I.3 --- Epstein-Barr Virus --- p.9 / Chapter I.3.1 --- Epidemiology --- p.9 / Chapter I.3.2 --- Virus and Genome Structure --- p.9 / Chapter I.3.3 --- Latent Infection --- p.11 / Chapter I.3.4 --- Latent Membrane Protein-1 --- p.12 / Chapter I.3.5 --- "EBV-Encoded, Small Non-polydenylated RNAs (EBERs)" --- p.13 / Chapter I.3.6 --- Disease Associated with EBV --- p.13 / Chapter I.3.7 --- EBV and PGL --- p.14 / Chapter I.4 --- Genetic Alterations --- p.15 / Chapter I.4.1 --- Background --- p.15 / Chapter I.4.2 --- Tumor Suppressor Genes (TSGs) --- p.16 / Chapter I.4.2.1 --- Origin and Structure of p53 Gene and Protein --- p.16 / Chapter I.4.2.2 --- Functions of p53 Gene --- p.18 / Chapter I.4.2.3 --- Inactivation Mechanisms of p53 --- p.21 / Chapter I.4.2.4 --- p53 Mutations and Protein Expression in NHLs --- p.24 / Chapter I.4.3 --- Oncogene --- p.25 / Chapter I.4.3.1 --- Bcl-2 --- p.25 / Chapter I.4.3.2 --- Other Oncogenes --- p.27 / Chapter II --- OBJECTIVES OF STUD Y --- p.30 / Chapter III --- ma terials and methods --- p.31 / Chapter III.1 --- Materials --- p.31 / Chapter III.2 --- Detection of EB V Latent Gene Product by In-situ Hybridization --- p.33 / Chapter III.2.1 --- Pretreatment of Paraffin-embedded Tissues and Apparatus --- p.33 / Chapter III.2.2 --- In-situ Hybridization of EBERs --- p.34 / Chapter III.3 --- Detection of p53 and bcl-2 and LMP-1 Protein Expression by Immunohisiochemistry --- p.35 / Chapter III.4 --- Microdissection of Formalin-fixed Paraffin-embedded Tissues --- p.37 / Chapter III.5 --- Extraction of Genomic DNA from Formalin-fixed Paraffin-embedded Tissues --- p.38 / Chapter III.5.1 --- Phenol / Chloroform Extraction --- p.38 / Chapter III.5.2 --- Commercially Available DNA Extraction Kit --- p.40 / Chapter III.6 --- Mutational Analysis p53 --- p.41 / Chapter III.6.1 --- Polymerase Chain Reaction - Single Strand Conformation Polymorphism (PCR-SSCP) Analysis --- p.41 / Chapter III.6.1.1 --- PCR primers --- p.41 / Chapter III.6.1.2 --- PCR Amplification ofp53 gene --- p.42 / Chapter III.6.1.3 --- Non-denaturing Polyacrylamide Gel Electrophoresis --- p.42 / Chapter III.6.2 --- DNA Sequencing Analysis --- p.44 / Chapter III.6.2.1 --- Purification of DNA from Shifts on Non-denaturing Gels --- p.44 / Chapter III.6.2.2 --- 5' end Labeling of Primer --- p.45 / Chapter III.6.2.3 --- Cycle Sequencing --- p.45 / Chapter III.6.2.4 --- Denaturing Gel Electrophoresis --- p.46 / Chapter III.7 --- Loss of Heterozygosity (LOH) Analysis on Chromosome 17p --- p.47 / Chapter III.7.1 --- Microsatellite Markers --- p.49 / Chapter III.7.2 --- PCR Amplification of DNA Fragments Containing Polymorphic Microsatellites --- p.49 / Chapter III.7.3 --- Denaturing Polyacrylamide Gel Electrophoresis --- p.50 / Chapter III.7.4 --- Determination of Allelic Abnormalities --- p.51 / Chapter III. 8 --- Statistical Analysis --- p.52 / Chapter IV --- results --- p.53 / Chapter IV.1 --- Association with Helicobactor Pylori (HP) --- p.53 / Chapter IV.2 --- Detection of EBERs by ISH --- p.53 / Chapter IV.3 --- Immunohistochemical Analysis --- p.54 / Chapter IV.3.1 --- Protein Expression of EBV LMP-1 --- p.54 / Chapter IV.3.2 --- Protein Expression of p53 --- p.54 / Chapter IV.3.3 --- Protein Expression of bcl-2 --- p.55 / Chapter IV.3.4 --- Correlation between p53 and bcl-2 Protein Expression --- p.55 / Chapter IV.4 --- Mutational Analysis of p53 --- p.56 / Chapter IV.5 --- LOH Analysis on Chromosome 17p --- p.57 / Chapter V --- DISCUSSION --- p.58 / Chapter V.1 --- Helicobactor Pylori Association --- p.58 / Chapter V.2 --- Association with EE V --- p.60 / Chapter V.3 --- Protein Expression of p53 and bcl-2 --- p.61 / Chapter V.3.1 --- p53 --- p.61 / Chapter V.3.2 --- Bcl-2 --- p.62 / Chapter V.3.3 --- Correlation between p53 and bcl-2 Expression --- p.63 / Chapter V.4 --- p53 Gene Molecular Analysis --- p.65 / Chapter V.5 --- Distribution of Mutations and Molecular Fingerprinting --- p.67 / Chapter V.6 --- Possible Role of p53 Mutation in EBV+ Gastric Lymphomas --- p.69 / ILLUSTRATIONS --- p.71 / references --- p.94

Chow Chit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-156). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Table of Contents --- p.vii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Malignant Lymphoma --- p.1 / Chapter 1.2 --- Non-Hodgkin's Lymphoma --- p.1 / Chapter 1.3 --- NK/T Cell Lymphoma --- p.2 / Chapter 1.3.1 --- General Features of NK/T Cell Lymphoma --- p.2 / Chapter 1.3.2 --- Histology of NK/T Cell Lymphoma --- p.3 / Chapter 1.3.3 --- Subtypes NK/T Cell Lymphoma --- p.4 / Chapter 1.3.4 --- Overview of NK/T Cell Lymphoma Cell Lines --- p.5 / Chapter 1.3.4.1 --- NK/T Cell Lymphoma Cell Lines-NK-92 and SNK-6 --- p.6 / Chapter 1.3.5 --- NK/T Cell Lymphoma and Interleukins --- p.8 / Chapter 1.4 --- Interleukin-2 and Interleukin-15 --- p.9 / Chapter 1.5 --- IL-2 and IL-15 Receptor --- p.10 / Chapter 1.6 --- Cellular Signaling Pathways Regulated by IL-2 and IL-15 --- p.11 / Chapter 1.6.1 --- JAK/STAT Pathway --- p.12 / Chapter 1.6.2 --- PI3K/Akt Pathway --- p.14 / Chapter 1.7 --- Epstein-Barr Virus: an Oncogenic Virus --- p.20 / Chapter 1.7.1 --- Overview of EBV --- p.20 / Chapter 1.7.2 --- Epidemiology --- p.20 / Chapter 1.7.3 --- Life Cycle of EBV --- p.21 / Chapter 1.7.4 --- Latency Infection of EBV --- p.21 / Chapter 1.7.5 --- Role of EBV latent genes in oncogenesis --- p.23 / Chapter 1.7.5.1 --- EBER1 and 2 --- p.23 / Chapter 1.7.5.2 --- EBNAs --- p.24 / Chapter 1.7.5.3 --- LMPs --- p.25 / Chapter 1.7.6 --- Lytic Cycle of EBV --- p.26 / Chapter 1.7.7 --- Signaling Pathways and EBV --- p.27 / Chapter Chapter 2: --- Aim of Study --- p.29 / Chapter Chapter 3: --- Materials and Methods --- p.31 / Chapter 3.1 --- IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines and patients --- p.31 / Chapter 3.1.1 --- Cell Lines Maintenance --- p.31 / Chapter 3.1.2 --- Patients --- p.32 / Chapter 3.2 --- "Assays of IL-2, IL-15 and IFN-γ in culture supernatants and patient sera" --- p.32 / Chapter 3.2.1 --- IL-2 ELISA --- p.32 / Chapter 3.2.2 --- IL-15 ELISA --- p.33 / Chapter 3.2.3 --- IFN-γ ELISA --- p.34 / Chapter 3.3 --- Effect of IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines --- p.35 / Chapter 3.3.1 --- Cell Growth and Viability Determination --- p.35 / Chapter 3.3.2 --- Apoptosis Assays on Interleukin-starved NK-92 cells --- p.35 / Chapter 3.3.2.1 --- DNA Laddering Analysis --- p.36 / Chapter 3.3.2.2 --- Cell Cycle and Apoptosis Determination by PI Staining --- p.37 / Chapter 3.3.2.3 --- Caspase 3 Activity Assay --- p.37 / Chapter 3.4 --- PI3K/Akt Pathway Study --- p.39 / Chapter 3.4.1 --- Determination of AKT1 Gene Amplification by Real-Time Quantitative PCR --- p.39 / Chapter 3.4.1.1 --- DNA Extraction for Real-Time Quantitative PCR --- p.39 / Chapter 3.4.1.2 --- AKT1 Real-Time Quantitative PCR --- p.40 / Chapter 3.4.2 --- Determination of Akt Expression --- p.41 / Chapter 3.4.2.1 --- Normal NK Cell Purification from Buffy Coat --- p.41 / Chapter 3.4.2.2 --- Determination of the Purity of Extracted NK Cells --- p.43 / Chapter 3.4.2.3 --- Interleukin Treatment of Normal NK Cells and NK-92 --- p.43 / Chapter 3.4.2.4 --- Protein Extraction and Western Blot Analysis --- p.43 / Chapter 3.4.3 --- Study of PI3 K/Akt pathway using PI3K inhibitor --- p.47 / Chapter 3.4.3.1 --- Cell Growth and Viability Assay --- p.47 / Chapter 3.4.3.2 --- Apoptosis Assay by DNA Laddering and Pi-Staining on NK-92 cells --- p.48 / Chapter 3.4.3.3 --- Determination of activated Akt after LY294002 and Wortmannin treatment --- p.48 / Chapter 3.5 --- Effect of IL-2 and IL-15 on the JAK/STAT pathway and PDK/Akt pathway of NK/T Cell Lymphoma Cell Lines --- p.49 / Chapter 3.5.1 --- Cell Treatment --- p.49 / Chapter 3.5.2 --- Study of JAK/STAT and PI3K/Akt Pathways by Western Blotting --- p.49 / Chapter 3.5.3 --- "Assays of IL-2, IL-15 and IFN-γ in the NK-92 Cell Culture Medium by ELISA" --- p.50 / Chapter 3.5.4 --- Determination of EBV Status after IL-2 and IL-15 Treatment --- p.51 / Chapter 3.5.4.1 --- RNA Extraction --- p.51 / Chapter 3.5.4.2 --- Reverse-transcriptase Reaction --- p.52 / Chapter 3.5.4.3 --- PCR for EBV-related Genes --- p.53 / Chapter 3.5.4.4 --- EBER-ISH --- p.54 / Chapter 3.5.4.5 --- Real-time Quantitative PCR for EBER1 --- p.56 / Chapter 3.5.4.6 --- Western Blot for LMP1 --- p.56 / Chapter 3.6 --- Statistical Analysis --- p.57 / Chapter Chapter 4: --- Results --- p.59 / Chapter 4.1.1 --- "IL-2, IL-15 and IFN-γ Levels in the Serum of Patients with NK/T Cell Lymphoma" --- p.59 / Chapter 4.1.2 --- IL-2 and IL-15 Level in Culture Supernatant of NK-92 --- p.59 / Chapter 4.1.3 --- IFN-γ induction in supernatant of NK-92 --- p.60 / Chapter 4.2 --- Effect of IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines --- p.61 / Chapter 4.2.1 --- Cell Growth and Viability --- p.61 / Chapter 4.2.2 --- Apoptosis Study of Interleukin-starved NK-92 --- p.62 / Chapter 4.2.2.1 --- DNA fragmentation and Cell Cycle studies --- p.62 / Chapter 4.2.2.2 --- Caspase 3 Activity in NK-92 --- p.62 / Chapter 4.3 --- Akt in NK-92 --- p.63 / Chapter 4.3.1 --- Confirmation ofAKTl Amplification in NK-92 --- p.63 / Chapter 4.3.2 --- Akt Protein Quantification in NK-92 cells --- p.63 / Chapter 4.3.3 --- Activated Akt and STAT proteins in IL-2 or IL-15 stimulated NK-92 and normal NK cells --- p.64 / Chapter 4.4. --- PI3K/Akt Pathway --- p.64 / Chapter 4.4.1 --- Phosphorylation of Components of the PI3K/Akt Pathway --- p.64 / Chapter 4.4.2. --- Role of PI3K/Akt Pathway in NK/T Cell Lymphoma Cell Lines --- p.65 / Chapter 4.4.2.1 --- Cell Growth and Viability Studies --- p.65 / Chapter 4.4.2.2 --- Apoptosis and Cell Cycle Arrest Induction by LY294002 in NK-92 --- p.66 / Chapter 4.4.2.3 --- Confirmation of the effect of LY294002 on the PDK/Akt pathway in NK-92 cells --- p.67 / Chapter 4.5 --- Phosphorylation of STAT family proteins --- p.67 / Chapter 4.6 --- Regulation of EBV-related genes in NK-92 --- p.68 / Chapter Chapter 5: --- Discussion --- p.71 / Chapter 5.1 --- Cytokine level in patient serum --- p.71 / Chapter 5.2 --- Source of IL-2 and IL-15 for NK-92 cells --- p.72 / Chapter 5.3 --- Induction of IFN-γ in NK-92 --- p.73 / Chapter 5.4 --- Role of IL-2 and IL-15 on NK/T cell lymphoma cell lines --- p.75 / Chapter 5.4.1 --- Cell Growth and Viability Maintenance by IL-2 and IL-15 --- p.75 / Chapter 5.4.2 --- Apoptosis induced by interleukin-starving in NK-92 --- p.75 / Chapter 5.5 --- Aberrant activation of signaling pathways by IL-2 or IL-15 in NK-92 --- p.77 / Chapter 5.5.1 --- Hypersensitivity of NK-92 cells to IL-2 or IL-15 --- p.77 / Chapter 5.5.2 --- PI3K/Akt pathway in NK/T cell lymphoma cell lines --- p.78 / Chapter 5.5.2.1 --- Confirmation of AKT1 amplification --- p.78 / Chapter 5.5.2.2 --- Constitutive activation of Akt in NK/T cell lymphoma cell lines --- p.79 / Chapter 5.5.2.3 --- Role of PI3K/Akt in NK/T cell lymphoma cell lines --- p.80 / Chapter 5.5.2.4 --- IL-2 and IL-15 induce differential sensitivity of NK-92 to LY294002 --- p.81 / Chapter 5.5.2.5 --- NK/T cell lymphoma cell lines are wortmannin-insensitive --- p.82 / Chapter 5.6 --- Jak/STAT pathway in NK/T cell lymphoma cell lines --- p.83 / Chapter 5.6.1 --- STAT3 and STAT5 were activated by both IL-2 and IL-15 --- p.83 / Chapter 5.6.2 --- STAT6 was activated in NK/T cell lymphoma cell lines --- p.84 / Chapter 5.6.3 --- "Differential regulation of STAT 1, STAT3 (Ser-727) and STAT6 in NK/T cell lymphoma cell lines" --- p.85 / Chapter 5.6 --- EBV gene regulation in NK-92 --- p.87 / Chapter Chapter 6: --- Conclusion --- p.91 / Tables --- p.93 / Figures --- p.102 / Reference --- p.128

Xu Xuequn. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 100-125). / Abstracts in English and Chinese. / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Epstein-Barr (EBV) Virus --- p.1 / Chapter 1.1.1 --- Virus Structure and Genome Structure --- p.1 / Chapter 1.1.2 --- Virus Types --- p.2 / Chapter 1.2 --- EBV Infection and malignancies --- p.3 / Chapter 1.2.1 --- In Vitro Infection --- p.3 / Chapter 1.2.2 --- Infection in the Natural Host --- p.8 / Chapter 1.2.3 --- Malignancies Associated with EBV --- p.11 / Chapter 1.3 --- T Cell-Mediated Immune Response to EBV --- p.16 / Chapter 1.3.1 --- The Pathway of Cell-Mediated Immune Response in Viral Infection --- p.16 / Chapter 1.3.2 --- Cell-Mediated Immune Response to EBV --- p.18 / Chapter 1.3.3 --- The Feature of CTLs Response to EBV --- p.20 / Chapter 1.4 --- CTLs to EBV Relevant MalignancieśؤApplications and Challenges --- p.21 / Chapter 1.5 --- HLA Polymorphisms and Strategy of Epitope-Based CTLs Therapy --- p.24 / Chapter 1.6 --- The Effect of HLA Polymorphism on EBV-Specific CTL Epitope Choice in Southern Chinese --- p.27 / Chapter 1.7 --- ELISPOT Assay 226}0ؤ Detection of CTLs Response --- p.32 / Chapter 1.8 --- Aim of This Study --- p.37 / Chapter Chapter 2: --- Material and Methods: --- p.39 / Chapter 2.1 --- Peptides --- p.39 / Chapter 2.2 --- PBMCs Preparations --- p.43 / Chapter 2.3 --- PBMC Counting and Cells Dilution --- p.43 / Chapter 2.4 --- Elispot Assay --- p.44 / Chapter 2.5 --- Counting the Spots --- p.45 / Chapter 2.6 --- Spots Forming Cells (SFC/106) and Positive Standard --- p.46 / Chapter Chapter 3: --- Results --- p.47 / Chapter 3.1 --- Validation of ELISPOT assay methodology --- p.47 / Chapter 3.2 --- CTLs Response to Each Epitope in the Population --- p.55 / Chapter 3.2.1 --- Positive Response to A11 Restricted and Mutant Epitopes in the Population --- p.55 / Chapter 3.2.2 --- Positive Frequencies of A2 Restricted Epitopes in the Population --- p.63 / Chapter 3.2.3 --- Positive Frequencies of Other HLA Allele Restriction Peptides --- p.70 / Chapter 3.3 --- CTLs Response Frequencies Categorized by Proteins --- p.74 / Chapter 3.3.1 --- "CTLs Response to LMP1, LMP2, EBNA1 Epitopes" --- p.74 / Chapter 3.3.2 --- "CTLs Response to EBNA2, EBNA-LP Epitopes, EBNA3 Epitopes" --- p.75 / Chapter 3.3.3 --- CTLs Response to LYTIC Epitopes --- p.79 / Chapter 3.4 --- Summary --- p.80 / Chapter Chapter 4: --- Discussion --- p.82 / Chapter 4.1 --- Discussion of A11 Restricted Epitopes --- p.82 / Chapter 4.2 --- Discussion of A2 Restricted Epitopes --- p.86 / Chapter 4.3 --- Discussion of Other HLA Restricted Epitopes --- p.89 / Chapter 4.4 --- "Discussion ofLMPl, LMP2, EBNA1 Epitopes" --- p.92 / Chapter 4.5 --- "Discussion of EBNA2, EBNA3, and EBNA-LP epitopes" --- p.96 / Chapter 4.6 --- Discussion of LYTIC Epitopes --- p.96 / Chapter 4.7 --- Discussion of Summary --- p.98 / Chapter Chapter 5 --- Conclusion --- p.99 / Chapter 6 --- Reference --- p.100 / Chapter 7 --- Appendix --- p.126 / Chapter 7.1 --- "Appendix 1, raw data of Elispot assay on CTLs response to EBV relevant epitopes m Hong Kong donors" --- p.126 / Chapter 7.2 --- "Appendix 2, frequencies from highest cell number wells of the peptides (SFC/106)" --- p.126 / Chapter 7.3 --- "Appendix 3, typical Elispot assay figure " --- p.126

1. Study on improving the diagnostic accuracy of treatment-naive nasopharyngeal carcinoma. / 2. Study on diagnostic accuracy of EBV-DNA on recurrent nasopharyngeal carcinoma. / 3. Studies on EBV-DNA as a screening tool for nasopharyngeal carcinoma. Part 1. To define the detection rate of NPC and the false-positive rate of IgA-VCA in an IgA-VCA-based screening problem, and to define the specificity of IgA-EA in IgA-VCA-positive screenees. Part 2. To define the specificity of EBV-DNA in IgA-VCA-positive screenees. Part 3. To define the sensitivity of IgA-EA, and EBV-DNA in IgA-positive NPC patients. / 4. Studies on pre-therapy prognostication of nasopharyngeal carcinoma Study Part 1. Objective. To assess the role of EBV-DNA in pre-therapy prognostication of early-stage NPC. / Background. The specific association between nasopharyngeal carcinoma (NPC) and the Epstein-Barr virus (EBV) had been exploited to develop a spectrum of EBV-antibodies-based blood markers. Among these markers, the Immunoglobulin A antibody against the viral capsid antigen (IgA-VCA) of the EBV has been the most popularly employed marker to assist diagnosis of NPC. There is however a relative paucity of data on the application of blood markers for screening, for detection of relapse, and for prognostification of patient cohorts managed in present-day therapy oncology protocols. Peripheral blood EBV-DNA, measured by quantitative polymerase chain reaction assay, is a newly-developed marker, and represents a prototype model of a nuclei acid-based, as opposed to antibody-based, EBV tumor marker for NPC. The present thesis describes the translation of this basic scientific advance into clinical applications, through several prospective and retrospective studies that address the diagnosis of treatment-naive NPC, the detection of recurrent NPC, the screening of individuals at risk of NPC, the pre-therapy prognostication for NPC to guide for choice of therapy. The role of integration of conventional markers and EBV-DNA in clinical applications was also examined. / Study Part 2. Objectives. To assess whether incorporation of EBV-DNA data to TNM staging improves prognostic discrimination of patients subsets within individual cancer stage, to assess if EBV-DNA is an independent prognostic factor for survival after ontological therapy. (Abstract shortened by UMI.) / Leung Sing-fai. / "February 2005." / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3695. / Thesis (M.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / School code: 1307.

A comprehensive immunohistochemical study was carried out to investigate the phenotypes and prevalence of intraepithelial lymphocytes in NPC samples semi-quantitatively. CD25+/FOXP3+ T-cells were highly prevalent in primary NPCs, suggesting the presence of the immunosuppressive Tregs in tumor microenvironments. The low abundance of CD4+ T-cells, and the positive correlation between FOXP3 and CD8 staining in NPC samples imply that CD8+FOXP3+ Tregs may be present and play role in the suppression of anti-tumor immune response in NPC patients. The involvement of chemokine in the migration of tumor-infiltrating lymphocytes was studied. Chemokine ligand 20 (CCL20) was overexpressed in all EBV-positive NPC cell lines and xenografts compared to EBV-negative NPC, and immortalized normal nasopharynx epithelial cell lines. The presence of CCL20 was also found in primary tumors but not in normal epithelium. Furthermore, the ability of LMP1 to upregulate CCL20 expression in epithelial cells indicates that EBV may induce the production of chemokine involved in lymphocyte migration. / Epstein-Barr virus (EBV) is invariably associated with the development of nasopharyngeal carcinoma (NPC). Although the association of EBV and cancer has been reported for about four decades, it is still not clear how EBV latent infection contributes to the transformation of nasopharyngeal epithelial cells. The aims of this study are to identify EBV-regulated cellular genes and pathways and to determine the potential role of EBV in the modulation of anti-tumor immune responses in NPC. / In summary, EBV plays critical roles in the development of NPC by regulation of multiple cellular genes and pathways such as the Notch signaling cascade, and modulation of anti-tumor immune responses through the induction of chemokine important in migration of immune cells. / Notch signaling pathway functions in diverse cellular processes such as proliferation, apoptosis, adhesion, and epithelial to mesenchymal transition. In the current study, aberrant expression of activated Notch1 receptor (NICD), Notch ligand (Jagged1), negative regulator of Notch ( NUMB) and Notch downstream effector (HEY1) was detected in NPC cell lines and xenografts. Overexpression of NICD, Jagged1 and HEY1 proteins was also commonly found in primary tumors of NPC. / Transfection of Jagged1 to normal nasopharynx epithelial cells resulted in increased cell proliferation. Moreover, EBERs, which is abundantly expressed in EBV-positive NPC tumors, was capable of inducing the expression of Jagged1 in epithelial cells. The current data shows that Notch signaling pathway is aberrantly activated by the deregulated expression of multiple Notch components in NPC. The induction of Jagged1 by EBERs also implies the potential role of EBV in the activation of Notch signaling cascade in NPC. / Using high-density oligonucleotide microarray, expression profiles of EBV-infected NPC cell lines, HK1+EBV and HONE1+EBV, and their uninfected counterparts, HK1 and HONE, were generated. From the microarray results, six EBV-upregulated (JDP2, IL8, ATP6V0E2L, PLAP, PIK3C2B and AKR1B10 ) and three EBV-downregulated genes (BACE2, PADI3 and MMP1) were identified in both HK1 and HONE1 cells upon EBV latent infection. One hundred and thirty-eight and seventy-six genes were also found to be differentially modulated by EBV in HK1 and HONE1 cells, respectively. This study shows that cellular genes involved in wide range of biological processes and cellular functions are differentially regulated by EBV, which suggest that EBV modulates multiple pathways and processes during NPC tumorigenesis. / Hui, Wai Ying. / Adviser: Kw Lo. / Source: Dissertation Abstracts International, Volume: 69-02, Section: B, page: 0806. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 166-204). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / School code: 1307.

Heung May-sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references. / Abstracts in English and Chinese. / Title Page --- p.i / Acknowledgement --- p.ii / Table of Contents --- p.iii / List of Abbreviations --- p.vi / List of Figures --- p.viii / List of Tables --- p.xii / Abstract --- p.xiv / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Epstein-Barr Virus --- p.1-1 / Chapter 1.1.1 --- Virus structure --- p.1-1 / Chapter 1.1.2 --- Genome structure --- p.1-1 / Chapter 1.1.3 --- Nomenclature for EBV open reading frames --- p.1-2 / Chapter 1.1.4 --- Biology of EBV --- p.1-2 / Chapter 1.1.5 --- EBV latency --- p.1-7 / Chapter 1.1.6 --- EBV latent gene promoters --- p.1-8 / Chapter 1.2 --- EBV Infection and Its Persisence --- p.1-9 / Chapter 1.3 --- DNA Methylation --- p.1-17 / Chapter 1.3.1 --- Aberrant CpG island methylation in cancer --- p.1-18 / Chapter 1.3.2 --- DNA methylation and EBV --- p.1-19 / Chapter 1.4 --- Demethylating Agents --- p.1-21 / Chapter 1.5 --- Aims of the Study --- p.1-23 / Chapter Chapter 2 --- EBV Latency Patterns / Chapter 2.1 --- Introduction --- p.2-1 / Chapter 2.2 --- Materials and Methods --- p.2-1 / Chapter 2.2.1 --- Cell line culture --- p.2-1 / Chapter 2.2.2 --- NPC biopsies culture --- p.2-2 / Chapter 2.2.3 --- RNA extraction --- p.2-3 / Chapter 2.2.4 --- RNA quantification --- p.2-4 / Chapter 2.2.5 --- Deoxyribonuclease I treatment for NPC biopsies --- p.2-5 / Chapter 2.2.6 --- Reverse transcriptase-polymerase chain reaction --- p.2-5 / Chapter 2.2.7 --- Gel Electrophoresis --- p.2-10 / Chapter 2.3 --- Results --- p.2-11 / Chapter 2.3.1 --- Burkitt' s lymphoma and lymphoblastoid cell lines --- p.2-11 / Chapter 2.3.2 --- Nasopharyngeal carcinoma biopsies --- p.2-11 / Chapter 2.4 --- Discussion --- p.2-19 / Chapter Chapter 3 --- Treatment with Demethylating Agents on Rael / Chapter 3.1 --- Introduction --- p.3-1 / Chapter 3.2 --- Materials and Methods --- p.3-2 / Chapter 3.2.1 --- Rael cell line culture --- p.3-2 / Chapter 3.2.2 --- Drug treatment --- p.3-2 / Chapter 3.2.3 --- Viability staining --- p.3-2 / Chapter 3.2.4 --- Statistical analysis --- p.3-3 / Chapter 3.2.5 --- RNA extraction and quantification --- p.3-3 / Chapter 3.2.6 --- RT-PCR and gel electrophoresis --- p.3-3 / Chapter 3.2.7 --- DIG oligonucleotide 3'-end labeling --- p.3-3 / Chapter 3.2.8 --- Southern blot --- p.3-10 / Chapter 3.3 --- Results --- p.3-13 / Chapter 3.3.1 --- 5-azacytidine --- p.3-13 / Chapter 3.3.2 --- 5-aza-2-deoxycytidine --- p.3-26 / Chapter 3.4 --- Discussion --- p.3-39 / Chapter Chapter 4 --- Treatment with Demethylating Agents on NPC Biopsies / Chapter 4.1 --- Introduction --- p.4-1 / Chapter 4.2 --- Materials and Methods --- p.4-2 / Chapter 4.2.1 --- NPC biopsy culture --- p.4-2 / Chapter 4.2.2 --- Drug treatment --- p.4-2 / Chapter 4.2.3 --- RNA extraction and quantification --- p.4-2 / Chapter 4.2.4 --- DNase I treatment for NPC biopsies --- p.4-2 / Chapter 4.2.5 --- RT-PCR and gel electrophoresis --- p.4-2 / Chapter 4.3 --- Results --- p.4-3 / Chapter 4.4 --- Discussion --- p.4-3 / Chapter Chapter 5 --- Conclusion --- p.5-1 / Reference --- p.R-1 / Appendix --- p.A-l

Orientador: Estela Kaminagakura / Coorientadora: Patrícia Pimentel de Barros / Banca: Ana Sueli Rodrigues Cavalcant / Banca: Luana Marotta Reis de Vasconcellos / Banca: Lia Mizobe Ono / Banca: Luciana Yamamoto de Almeida / Resumo: A papilomatose laríngea é uma neoplasia benigna causada pelo papilomavírus humano (HPV), sendo os tipos 6 e 11 os mais comuns, e que ocorre em dois grupos etários, juvenil e adulto. A possível coinfecção viral tem sido sugerida em lesões de cabeça e pescoço; nesse sentido, o Epstein Barr vírus (EBV), que também apresenta tropismo por células epiteliais vem sendo estudado neste grupo de lesões. Os objetivos deste estudo foram genotipar os HPVs, investigar a presença de EBV-DNA por PCR e EBV-RNA por hibridização in situ. Além disso, associar a presença de EBV com a imunoexpressão de CD21, os resultados obtidos com a escala laringoscópica de Derkay et al. (1998) e com os dados clinicopatológicos. Oitenta casos de papilomatose laríngea, juvenil (n=36) e adulta (n=44), foram retrospectivamente analisados e subdivididos em grupos de menor e maior severidade, baseando-se na escala de Derkay. Todas as amostras foram HPV posivitas, com 49 casos HPV 6, 26 casos HPV 11, 4 casos HPV 6 e 11, e 1 caso HPV 16. A presença de EBV-DNA foi detectada em 9 amostras, entretanto EBV-RNA não foi não foi identificado em nenhuma amostra. Assim como a presença do EBV-DNA, a imunoexpressão de CD21 não se associou estatisticamente com quaisquer variáveis. A presença de HPV 6 foi mais comum em PLA e, o HPV 11 foi mais comum (p=0,02) e maior em casos de maior severidade (p=0,04), no grupo juvenil. A presença do EBV provavelmente não desempenha papel importante na progressão/severidade desta patologia. / Doutor

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 3 uppsatser.

Zugl.: Berlin, Freie Univ., Diss., 2009