by Li Ho Kin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 213-245). / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.iii / ABSTRACT --- p.viii / TABLE OF CONTENTS --- p.xi / Chapter CHAPTER 1: --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Introduction --- p.2 / Chapter 1.2 --- Effector Cells Mediating Anti-Tumour Immunity --- p.5 / Chapter 1.2.1 --- Cytotoxic T Lymphocytes --- p.6 / Chapter 1.2.2 --- Natural Killer Cells --- p.7 / Chapter 1.2.3 --- Lymphokine-Activated Killer Cells --- p.9 / Chapter 1.2.4 --- Macrophages --- p.10 / Chapter 1.2.5 --- Tumour-Infiltrating Lymphocytes --- p.12 / Chapter 1.3 --- Current Modalities for Cancer Treatment --- p.13 / Chapter 1.3.1 --- Surgery --- p.13 / Chapter 1.3.2 --- Radiation Therapy --- p.14 / Chapter 1.3.3 --- Chemotherapy --- p.14 / Chapter 1.3.4 --- Biologic Therapy --- p.15 / Chapter 1.4 --- Biological Response Modifiers as Cancer Therapeutics --- p.17 / Chapter 1.4.1 --- Monoclonal Antibodies --- p.20 / Chapter 1.4.2 --- Differentiating Agents --- p.21 / Chapter 1.4.3 --- Cytokines --- p.24 / Chapter 1.4.4 --- Bioactive Plant Polysaccharides --- p.30 / Chapter 1.5 --- Bacterial Polysaccharides as Potential Immunomodulators and Anti-Tumour Agents --- p.36 / Chapter 1.5.1 --- General Properties of Bacterial Polysaccharides --- p.36 / Chapter 1.5.2 --- Structure and Function of Bacterial Polysaccharides --- p.37 / Chapter 1.5.3 --- Immunomodulatory and Anti-Tumour Activities of Bacterial Polysaccharides --- p.39 / Chapter 1.5.4 --- General Properties of Klebsiella K7 Capsular Antigen --- p.41 / Chapter 1.6 --- Aims and Scopes of the Investigation --- p.44 / Chapter CHAPTER 2: --- MATERIALS AND METHODS --- p.46 / Chapter 2.1 --- Materials --- p.47 / Chapter 2.1.1 --- Animals --- p.47 / Chapter 2.1.2 --- Klebsiella pneumoniae Serotype7 --- p.47 / Chapter 2.1.3 --- Agar Medium for Cultivation of Klebsiella pneumoniae Serotype7 --- p.47 / Chapter 2.1.4 --- Cell Lines --- p.47 / Chapter 2.1.5 --- "Buffers, Culture Medium and Other Reagents" --- p.49 / Chapter 2.1.6 --- Antibodies --- p.54 / Chapter 2.1.7 --- Radioisotopes --- p.55 / Chapter 2.1.8 --- Recombinant Cytokines --- p.55 / Chapter 2.1.9 --- Oligonucleotide Primers and Internal Probes --- p.56 / Chapter 2.1.10 --- Reagents and Solutions for Gene Expression Analysis --- p.59 / Chapter 2.2 --- Methods --- p.64 / Chapter 2.2.1 --- "Extraction, Purification and Characterization of Klebsiella K7 Capsular Antigen" --- p.54 / Chapter 2.2.1.1 --- Extraction and Purification of K7 Capsular Antigen --- p.64 / Chapter 2.2.1.2 --- Gel Filtration of K7 Capsular Antigen --- p.65 / Chapter 2.2.1.3 --- Characterization of K7 Capsular Antigen --- p.67 / Chapter 2.2.1.4 --- Determination of the Bio-Toxicity of K7 Capsular Antigen --- p.67 / Chapter 2.2.1.5 --- In Vitro Cytotoxicity of K7 Capsular Antigen on Splenocytes --- p.53 / Chapter 2.2.2 --- Assay for the Hematopoietic and Mitogenic Activities of Klebsiella K7 Capsular Antigen --- p.69 / Chapter 2.2.2.1 --- Isolation and Preparation of Cells --- p.69 / Chapter 2.2.2.2 --- In Vitro Lymphocyte Transformation Assay --- p.70 / Chapter 2.2.2.3 --- In Vitro Assay of Thymocyte Proliferation --- p.70 / Chapter 2.2.2.4 --- Depleting Mouse T Cells by Anti-Thy-1.2 Antibody Plus Complement Treatment --- p.71 / Chapter 2.2.2.5 --- Depleting Mouse B Cells by Anti-mouse B Cell Antibody Plus Complement Treatment --- p.71 / Chapter 2.2.2.6 --- Depleting Macrophages from Spleen Cell Suspension --- p.71 / Chapter 2.2.2.7 --- Flow Cytometric Analysis of Different Cell Populations from Splenocytes --- p.72 / Chapter 2.2.2.8 --- In Vitro Assay of rmIL-3-Stimulated Proliferation of Murine Bone Marrow Cells --- p.73 / Chapter 2.2.2.9 --- In Vitro Cytotoxicity of K7 Capsular Antigen on Bone Marrow Cells --- p.73 / Chapter 2.2.2.10 --- Colony Assay of Murine Bone Marrow Cells --- p.73 / Chapter 2.2.2.11 --- Assay of Differentiation of Murine Bone Marrow Cells --- p.74 / Chapter 2.2.3 --- Assay for Macrophage Activating Activities of Klebsiella K7 Capsular Antigen --- p.75 / Chapter 2.2.3.1 --- Preparation of Murine Peritoneal Exudate Cells (PEC) --- p.75 / Chapter 2.2.3.2 --- Assay of Phagocytic Activity of Peritoneal Macrophages --- p.75 / Chapter 2.2.3.3 --- In Vitro Macrophage-Mediated Cytostatic Activity --- p.76 / Chapter 2.2.3.4 --- Nitric Oxide (NO) Production of Peritoneal Macrophages --- p.76 / Chapter 2.2.3.5 --- In Vivo Migration of Macrophages --- p.77 / Chapter 2.2.3.6 --- Tumour Necrosis Factor (TNF) Production by Peritoneal Macrophages --- p.77 / Chapter 2.2.3.7 --- TNF Bioassay --- p.78 / Chapter 2.2.3.8 --- Gene Expression Analysis in Peritoneal Macrophages --- p.78 / Chapter 2.2.3.8.1 --- Preparation of Cell Lysate --- p.78 / Chapter 2.2.3.8.2 --- RNA Isolation --- p.79 / Chapter 2.2.3.8.3 --- Reverse Transcription of RNA --- p.80 / Chapter 2.2.3.8.4 --- Polymerase Chain Reaction (PCR) --- p.80 / Chapter 2.2.3.8.5 --- Agarose Gel Electrophoresis --- p.82 / Chapter 2.2.3.8.6 --- 3' End Labelling of Oligonucleotide Probes --- p.82 / Chapter 2.2.3.8.7 --- Dot Blot Hybridization --- p.83 / Chapter 2.2.3.8.8 --- DIG Chemiluminescent Detection --- p.83 / Chapter 2.2.4 --- Assay for Anti-Tumour Activities of Klebsiella K7 Capsular Antigen --- p.85 / Chapter 2.2.4.1 --- Assay of Tumour Cell Proliferation --- p.85 / Chapter 2.2.4.2 --- Assay of Anti-Tumour Activity In Vivo --- p.85 / Chapter 2.2.4.3 --- Assay of Lymphokine-Activated Killer Cell Activity --- p.85 / Chapter 2.2.4.4 --- Assay of Natural Killer Cell Activity --- p.87 / Chapter 2.2.4.5 --- Assay for Differentiation-Associated Characteristics of Myeloid Leukemia Cells --- p.88 / Chapter 2.2.4.5.1 --- Determination of the Viability of Myeloid Leukemia JCS cells --- p.88 / Chapter 2.2.4.5.2 --- Assessment of Cell Morphology --- p.88 / Chapter 2.2.4.5.3 --- Surface Antigen Immunophenotyping --- p.88 / Chapter 2.2.4.5.4 --- Assay of Non-specific Esterase Activity --- p.89 / Chapter 2.2.4.5.5 --- Assay of Phagocytosis --- p.90 / Chapter 2.2.5 --- Statistical Analysis --- p.90 / Chapter CHAPTER 3: --- "EXTRACTION, PURIFICATION & CHARACTERIZATION OF KLEBSIELLA K7 CAPSULAR ANTIGEN" --- p.91 / Chapter 3.1 --- Introduction --- p.92 / Chapter 3.2 --- Results --- p.94 / Chapter 3.2.1 --- Extraction and Purification of K7 Capsular Antigen from Klebsiella pneumoniae Serotype7 --- p.94 / Chapter 3.2.2 --- Gel Filtration Chromatography of K7 Capsular Antigen --- p.94 / Chapter 3.2.3 --- Characterization of K7 Capsular Antigen --- p.97 / Chapter 3.2.4 --- Determination of Bio-toxicity and Cellular Toxicity of K7 Capsular Antigen --- p.97 / Chapter 3.3 --- Discussion --- p.101 / Chapter CHAPTER 4: --- HEMATOPOIETIC AND MITOGENIC ACTIVITIES OF KLEBSIELLA K7 CAPSULAR ANTIGEN --- p.103 / Chapter 4.1 --- Introduction --- p.104 / Chapter 4.2 --- Results --- p.107 / Chapter 4.2.1 --- Effect of K7 Capsular Antigen on the In Vitro Proliferation of Murine Bone Marrow Cells --- p.107 / Chapter 4.2.2 --- Effect of K7 Capsular Antigen on rmIL-3-Stimulated Proliferation of Murine Bone Marrow Cells --- p.107 / Chapter 4.2.3 --- Effect of K7 Capsular Antigen on the In Vitro Differentiation of Murine Bone Marrow Cells --- p.112 / Chapter 4.2.4 --- Effect of K7 Capsular Antigen on the In Vitro Murine Bone Marrow Colony Formation --- p.112 / Chapter 4.2.5 --- Effect of K7 Capsular Antigen on the Viability of Splenocytes In Vitro --- p.117 / Chapter 4.2.6 --- In Vitro Mitogenic Effect of K7 Capsular Antigen on Splenocytes and Thymocytes --- p.117 / Chapter 4.2.7 --- Characterization of the Lymphocyte Population(s) Responding to K7 Capsular Antigen --- p.121 / Chapter 4.2.8 --- Effect of Polymyxin B Sulphate on the Mitogenic Activity of K7Capsular Antigen --- p.125 / Chapter 4.2.9 --- In Vivo Mitogenic Activity of K7 Capsular Antigen --- p.125 / Chapter 4.2.10 --- Flow Cytometric Analysis of Splenocytes from K7 Capsular Antigen-Treated Mice --- p.128 / Chapter 4.2.11 --- In Vitro Co-mitogenic Activity of K7 Capsular Antigen --- p.128 / Chapter 4.3 --- Discussion --- p.132 / Chapter CHAPTER 5: --- ACTIVATION OF MACROPHAGES BY KLEBSIELLA K7CAPSULAR ANTIGEN --- p.136 / Chapter 5.1 --- Introduction --- p.137 / Chapter 5.2 --- Results --- p.140 / Chapter 5.2.1 --- Effect of K7 Capsular Antigen on the Phagocytic Activity of Macrophages In Vitro --- p.140 / Chapter 5.2.2 --- Effect of K7 Capsular Antigen on the In Vivo Migration of Macrophages --- p.140 / Chapter 5.2.3 --- Effect of K7 Capsular Antigen on the In Vitro Cytostatic Activity of Picolinic Acid (PLA)-Activated Macrophages --- p.144 / Chapter 5.2.4 --- Effect of K7 Capsular Antigen on Nitric Oxide (NO) Production by Macrophages In Vitro --- p.144 / Chapter 5.2.5 --- Effect of K7 Capsular Antigen on the In Vitro Production of Tumour Necrosis Factor (TNF) by Macrophages --- p.148 / Chapter 5.2.6 --- Effect of K7 Capsular Antigen on the In Vitro Induction of Gene Expression in Macrophages --- p.148 / Chapter 5.3 --- Discussion --- p.166 / Chapter CHAPTER 6: --- ANTI-TUMOUR ACTIVITIES OF KLEBSIELLA K7 CAPSULAR ANTIGEN --- p.171 / Chapter 6.1 --- Introduction --- p.172 / Chapter 6.2 --- Results --- p.174 / Chapter 6.2.1 --- Effect of K7 Capsular Antigen In Vitro Growth of Various Tumour Cell Lines --- p.174 / Chapter 6.2.2 --- Effect of K7 Capsular Antigen on the In Vitro Growth of EAT Cells --- p.174 / Chapter 6.2.3 --- Effect of K7 Capsular Antigen on the Activation of LAK cells --- p.185 / Chapter 6.2.4 --- Effect of K7 Capsular Antigen on the Activation of NK Cells --- p.185 / Chapter 6.2.5 --- Effect of K7 Capsular Antigen on the Induction of Monocytic Differentiation of the Murine Myeloid Leukemia JCS Cells --- p.185 / Chapter 6.2.5.1 --- Effect of K7 Capsular Antigen on the Induction of Morphological Changes of JCS Cells --- p.188 / Chapter 6.2.5.2 --- Effect of K7 Capsular Antigen on the Expression of Macrophage Differentiation Antigen on Leukemia JCS Cells --- p.192 / Chapter 6.2.5.3 --- Effect of K7 Capsular Antigen on the Induction of Non-specific Esterase Activity in Leukemia JCS Cells --- p.192 / Chapter 6.2.5.4 --- Effect of K7 Capsular Antigen on the Stimulation of Phagocytic Activity in Leukemia JCS Cells --- p.192 / Chapter 6.3 --- Discussion --- p.198 / Chapter CHAPTER 7: --- CONCLUSIONS & FUTURE PERSPECTIVES --- p.202 / REFERENCES --- p.213
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_321862 |
| Date | January 1997 |
| Contributors | Li Ho Kin., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvii, 245 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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