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

by Ho Cheong Yip. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 152-164). / ACKNOWLEDGMENTS --- p.I / ABBREVIATIONS --- p.II / ABSTRACT --- p.VI / CHAPTER / Chapter 1 --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Immunomodulators of biological origin --- p.1 / Chapter 1.2 --- Effector cells involved in anti-tumor immunity --- p.3 / Chapter 1.2.1 --- Cytotoxic T lymphocytes --- p.3 / Chapter 1.2.2 --- Macrophages --- p.4 / Chapter 1.2.3 --- Natural killer cells --- p.5 / Chapter 1.2.4 --- Lymphokine-activated killer cells --- p.7 / Chapter 1.2.5 --- Tumor-infiltrating lymphocytes --- p.9 / Chapter 1.3 --- Cytokines involved in immunomodulation and anti-tumor immunity --- p.11 / Chapter 1.3.1 --- Interleukin-1 --- p.11 / Chapter 1.3.2 --- Interleukin-2 --- p.13 / Chapter 1.3.3 --- Interleukin-3 --- p.14 / Chapter 1.3.4 --- Tumor necrosis factor-alpha --- p.15 / Chapter 1.3.5 --- Interferons --- p.18 / Chapter 1.4 --- Carbohydrates used as potential immunopotentiating agents --- p.19 / Chapter 1.5 --- General properties of K1 capsular antigen --- p.20 / Chapter 2 --- AIM AND SCOPE OF THIS DISSERTATION --- p.22 / Chapter 3 --- MATERIALS AND METHODS --- p.24 / Chapter 3.1 --- Materials --- p.24 / Chapter 3.2 --- Methods --- p.33 / Chapter (I) --- "Extraction, purification and characterization of K1 capsular antigen from Klebsiella pneumoniae" / Chapter 3.2.1 --- Extraction and purification of Kl capsular antigen --- p.33 / Chapter 3.2.2 --- Gel filtration of K1 capsular antigen --- p.35 / Chapter 3.2.3 --- Characterization of K1 capsular antigen --- p.35 / Chapter 3.2.3.1 --- Determination of carbohydrate and protein contents --- p.35 / Chapter 3.2.3.2 --- Determination of uronic acid content --- p.35 / Chapter 3.2.4 --- Determination of bio-toxicity of K1 capsular antigen --- p.35 / Chapter 3.2.5 --- Treatment of K1 capsular antigen with sodium periodate --- p.36 / Chapter 3.2.6 --- Treatment of K1 capsular antigen with sodium hydroxide followed by acetic acid neutralization --- p.36 / Chapter (II) --- Isolation and preparation of cells / Chapter 3.2.7 --- Murine splenocytes --- p.37 / Chapter 3.2.8 --- Removal of red blood cells and dead cells using Ficoll-paque gradient method --- p.37 / Chapter 3.2.9 --- Murine thymocytes --- p.37 / Chapter 3.2.10 --- Murine peritoneal exudate cells (PEC) --- p.38 / Chapter 3.2.11 --- Bone marrow cells --- p.38 / Chapter (III) --- Assays of immunomodulatory activities of K1 capsular antigen on lymphocytes / Chapter 3.2.12 --- In vitro mitogenic assay --- p.39 / Chapter 3.2.13 --- In vivo mitogenic assay --- p.39 / Chapter 3.2.14 --- "In vitro co-mitogenic assay of K1 capsular antigen with Polymyxin B sulfate, LPS or Con A" --- p.40 / Chapter 3.2.15 --- In vitro mitogenic effect of K1 capsular antigen on lymphocyte sub-populations --- p.40 / Chapter 3.2.16 --- Assay of murine interleukin-2 --- p.41 / Chapter 3.2.17 --- Assay of murine interleukin-1 --- p.41 / Chapter (IV) --- Assays of immunomodulatory activities of K1 capsular antigen on macrophages and bone marrow cells / Chapter 3.2.18 --- In vivo migration of macrophages --- p.42 / Chapter 3.2.19 --- Assay of interleukin-1 produced from murine macrophages --- p.42 / Chapter 3.2.20 --- Assay of nitric oxide produced from murine macrophages --- p.43 / Chapter 3.2.21 --- Assay of proliferation of murine bone marrow cells --- p.44 / Chapter 3.2.22 --- Assay of differentiation of murine bone marrow cells --- p.44 / Chapter (V) --- Assays of anti-tumor activities of K1 capsular antigen / Chapter 3.2.23 --- In vitro cytostatic effect of K1 capsular antigen on murine tumor cell lines --- p.45 / Chapter 3.2.24 --- In vivo anti-tumor activities of K1 capsular antigen --- p.45 / Chapter 3.2.25 --- In vitro stimulation of TNF-α-like factor release from thioglycollate-elicited murine peritoneal macrophages by K1 capsular antigen --- p.46 / Chapter 3.2.26 --- Effects of K1 capsular antigen on TNF-α production in normal mice in vivo --- p.47 / Chapter 3.2.27 --- Effects of K1 capsular antigen on TNF-α production as well as EAT growth in vivo --- p.48 / Chapter 3.2.28 --- In vitro induction of macrophage-mediated cytostasis on tumor cells --- p.48 / Chapter 3.2.29 --- In vitro induction of macrophage-mediated cytolysis on tumor cells --- p.49 / Chapter 3.2.30 --- In vivo induction of alloreactive cytotoxic T cells --- p.49 / Chapter 3.2.31 --- In vitro induction of lymphokine-activated killer cell activity --- p.50 / Chapter 3.2.32 --- In vivo induction of lymphokine-activated killer cell activity --- p.51 / Chapter 3.2.33 --- In vivo induction of lymphokine-activated killer cell activity in tumor-bearing mice --- p.52 / Chapter 3.2.34 --- In vivo induction of lymphokine-activated killer cell activity in tumor-bearing mice with in vitro culture --- p.52 / Chapter 3.2.35 --- Phenotypic characterization of LAK cells by flow cytometry --- p.53 / Chapter 3.2.36 --- Winn-type tumor-inhibition of LAK cells --- p.54 / Chapter 3.2.37 --- Adoptive transfer of LAK cells to tumor-bearing mice --- p.54 / Chapter 3.2.38 --- In vivo stimulation of tumor-infiltrating lymphocytes --- p.55 / Chapter 3.2.39 --- Statistical analysis --- p.57 / Chapter 4 --- "EXTRACTION, PURIFICATION AND CHARACTERIZATION OF K1 CAPSULAR ANTIGEN FROM KLEBSIELLA PNEUMONIAE" / Introduction --- p.58 / Results --- p.59 / Chapter 4.1 --- Extraction and purification of Kl capsular antigen from Klebsiella pneumoniae --- p.59 / Chapter 4.2 --- Gel filtration of Kl capsular antigen --- p.59 / Chapter 4.3 --- Characterization of Kl capsular antigen --- p.62 / Chapter 4.4 --- Determination of toxicity (LC50) of Kl capsular antigen by brine shrimp assay --- p.62 / Discussion --- p.62 / Chapter 5 --- IMMUNOMODULATORY ACTIVITIES OF K1 CAPSULAR POLYSACCHARIDE ANTIGEN FROM KLEBSIELLA PNEUMONIAE / Introduction --- p.67 / Results --- p.69 / Chapter 5.1 --- Mitogenic activity of Kl capsular antigen on murine lymphocytes in vitro --- p.69 / Chapter 5.2 --- Mitogenic activity of K1 capsular antigen on murine lymphocytes in vivo --- p.69 / Chapter 5.3 --- Evidences to support the mitogenic activity of K1 capsular antigen is due to its polysaccharide structure rather than due to the contamination by LPS --- p.74 / Chapter 5.4 --- Effect of Kl capsular antigen on IL-2 production from murine lymphocytes in vitro --- p.79 / Chapter 5.5 --- Effect of K1 capsular antigen on IL-1 -like factor production from murine thymocytes in vitro --- p.79 / Chapter 5.6 --- Immunopotentiating activities of Kl capsular antigen on macrophages --- p.85 / Chapter 5.6.1 --- In vivo migration of macrophages in K1-treated mice --- p.85 / Chapter 5.6.2 --- Effect ofKl capsular antigen on the macrophage EL-1-like factor production in vitro --- p.85 / Chapter 5.6.3 --- Effect ofKl capsular antigen on the macrophage nitric oxide (NO) production in vitro --- p.85 / Chapter 5.7 --- Effects of Kl capsular antigen on the proliferation and differentiation of murine bone marrow cells --- p.89 / Discussion ´ب --- p.94 / Chapter 6 --- ANTI-TUMOR ACTIVITIES OF K1 CAPSULAR POLYSACCHARIDE ANTIGEN FROM KLEBSIELLA PNEUMONIAE / Introduction --- p.100 / Results --- p.102 / Chapter 6.1 --- In vitro cytostatic effects of Kl capsular antigen on murine tumor cell lines --- p.102 / Chapter 6.2 --- In vivo anti-tumor activities of Kl capsular antigen --- p.102 / Chapter 6.3 --- Effects of Kl capsular antigen on TNF-α production and on EAT growth in vivo --- p.110 / Chapter 6.4 --- In vitro induction of macrophage-mediated cytostatic effect on tumor cells by K1 capsular antigen --- p.110 / Chapter 6.5 --- In vitro induction of macrophage-mediated cytolytic effect on tumor cells by K1 capsular antigen --- p.115 / Chapter 6.6 --- Effect of Kl capsular antigen on the activation of alloreactive cytotoxic T cells --- p.115 / Chapter 6.7 --- Effect of Kl capsular antigen on the activation of lymphokine- activated killer (LAK) cells in vitro and in vivo --- p.115 / Chapter 6.8 --- Phenotypic characterization of LAK cells by flow cytometry --- p.123 / Chapter 6.9 --- Winn-type tumor inhibition assay of LAK cells --- p.126 / Chapter 6.10 --- Adoptive transfer of LAK cells to tumor-bearing mice --- p.132 / Chapter 6.11 --- Effect ofKl capsular antigen on the activation of tumor- infiltrating lymphocytes (TILs) in tumor-bearing mice --- p.132 / Discussion --- p.136 / Chapter 7 --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.146 / BIBLIOGRAPHY --- p.152

by Tang Yan Chi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 156-168). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABBREVIATIONS --- p.II / ABSTRACT --- p.V / ABSTRACT (CHINESE) --- p.VII / TABLE OF CONTENT --- p.1 / GENERAL INTRODUCTION --- p.4 / Chapter 1.1. --- Immunotherapy in Cancer --- p.4 / Chapter 1.1.2. --- Cytokines used in Immunotherapy in Cancer --- p.5 / Chapter 1.1.2.1. --- Interferons (IFNs) --- p.6 / Chapter 1.1.2.2. --- interleukins (ILs) --- p.11 / Chapter 1.1.2.3. --- Tumor Necrosis Factor-Alpha (TNF-α) --- p.14 / Chapter 1.2. --- Immunomodulatory and Anti-tumor Activities of Plant Polysaccharides --- p.17 / Chapter 1.3. --- Previous Studies on the Capsular Polysaccharides (CPS) from Klebsiella pneumoniae --- p.21 / Chapter 1.4. --- Activation of TNF-α Production from Macrophages by Bacterial Polysaccharides --- p.26 / Chapter 1.5. --- Chemotherapy of Cancer --- p.29 / Chapter 1.5.1. --- Types of Chemotherapeutic Drugs --- p.29 / Chapter 1.5.1.1. --- Alkylating Agents --- p.29 / Chapter 1.5.1.2. --- Plant Alkaloids --- p.31 / Chapter 1.5.1.3. --- Anti-metabolites --- p.32 / Chapter 1. 5.1.4. --- Antitumor Antibiotics --- p.32 / Chapter 1.5.2. --- Chemotherapeutic Drugs in this Project --- p.34 / Chapter 1.5.2.1. --- actinomycin D (ACT D) --- p.34 / Chapter 1.5.2.2. --- Cyclophosphamide (CY) --- p.35 / Chapter 1.5.2.3. --- Doxorubicin (Dox) --- p.37 / Chapter CHAPTER TWO --- AIM AND SCOPE OF THIS DISSERTATION --- p.40 / Chapter CHAPTER THREE --- MATERIALS AND METHODS --- p.42 / Chapter 3.1 --- MATERIALS --- p.42 / Chapter 3.1.1. --- Animals --- p.42 / Chapter 3.1.2. --- Klebsiella pneumoniae Serotype 1 and Serotype 24 --- p.42 / Chapter 3.1.3. --- Cell lines --- p.43 / Chapter 3.1.4. --- "Buffers, culture media and chemicals" --- p.43 / "POLYCLONAL RABBIT ANTI-ACTIVE ERK1/2, ANTI-ACTIVE JNK 1/2 AND ANTI-ACTIVE P38WERE FROM PROMEGA. POLYCLONAL RABBIT ANTI-TOTAL JNK1, ANTI-TOTAL P38WERE FROM SANTA CRUZ. MONOCLONAL ANTI-MOUSE TOTAL ERK1 AND POLYCLONAL ANTI-RABBIT TOTAL ERK2 WERE PURCHASED FROM ZYMED" --- p.50 / Chapter 3.2. --- METHODS --- p.51 / Chapter 3.2.1. --- "Extraction, Purification and Characterization of K1 and K24 Capsular Polysaccharides (CPS) from Klebsiella pneumoniae" --- p.51 / Chapter 3.2.1.1. --- Extraction and Purification of K1 and K24 Klebsiella Capsular Polysaccharides (CPS) --- p.51 / Chapter 3.2.1.2. --- Gel filtration of K1 and K24 Klebsiella Capsular Polysaccharides --- p.52 / Chapter 3.2.1.3. --- Characterization of K1 and K24 Capsular Polysaccharides --- p.52 / Chapter 3.2.3. --- Assay for Macrophage Activating Activities of Klebsiella Capsular Polysaccharides --- p.55 / Chapter 3.2.3.1. --- Tumour Necrosis Factor-α (TNF-α) Production by Peritoneal Macrophages --- p.55 / Chapter 3.2.3.2. --- Effect of Capsular Polysaccharides on the activities of MAPK family in murine macrophages --- p.56 / Chapter 3.2.4. --- Assay for Anti-Tumor Activities of Klebsiella Capsular Polysaccharides --- p.63 / Chapter 3.2.4.1. --- Assay of in vivo Suppression of EAT growth by K1 and K24 Capsular Polysaccharides --- p.63 / Chapter 3.2.4.2. --- Assay of the Effect of CPS on the Survival of EAT-Bearing Mice --- p.64 / Chapter 3.2.5. --- "Assay for Anti-Tumor Activities of Combined Treatment of Klebsiella Capsular Polysaccharides and Chemotherapeutic Drugs (Actinomycin D (Act D), Cyclophosphamide (CY) and Doxorubicin (Dox)" --- p.65 / Chapter 3.2.5.1. --- "Assay of the Cytotoxic Effect of Chemotherapeutic Drugs： Actinomycin D (Act D), Cyclophosphamide (CY) and Doxorubicin (Dox), on wehi-164 cell line" --- p.65 / Chapter 3.2.5.2. --- "Assay of the Combined Treatment of the CPS and Chemotherapeutic Drugs (Act D, CY and Dox) on WEHI-164 cell line in vitro" --- p.66 / Chapter 3.2.5.3. --- Assay for Anti-Tumor Effect of Chemotherapeutic Drugs (Act D，CY and Dox) on EAT-bearing Mice in vivo --- p.67 / Chapter 3.2.5.4. --- "Assay of the Combined Treatment of the CPS and Chemotherapeutic Drugs (Act D, CY and Dox) on EAT-bearing Mice in vivo" --- p.68 / Chapter 3.2.5.5. --- "Assay of the Combined Treatment of the CPS and Chemotherapeutic Drugs (Act D, CY and Dox) on the survival of eat-bearing mice in vivo" --- p.72 / Chapter CHAPTER FOUR --- "EXTRACTION, PURIFICATION & CHARACTERIZATION OF KLEBSIELLA K1 & K24 CAPSULAR POLYSACCHARIDES" --- p.76 / Chapter 4.1. --- Extraction and Purification of K1 and K24 Capsular Polysaccharides from Klebsiella pneumoniae Serotype 1 and Serotype 24 --- p.76 / Chapter 4.2. --- Gel Filtration Chromatography of K1 and K24 Capsular Polysaccharides --- p.77 / Chapter 4.3. --- Characterization of K1 and K24 Capsular Polysaccharides --- p.81 / Chapter CHAPTER FIVE --- EFFECT OF THE K1 & K24 CAPSULAR POLYSACCHARIDES ON THE MACROPHAGE ACTIVITIES --- p.83 / Chapter 5.1. --- Effect ofK1 and K24 Capsular Polysaccharides on the in vitro induction of TNF- α Production of Murine Peritoneal Macrophages --- p.83 / Chapter 5.2. --- Effect of Capsular Polysaccharides on the activities of MAPK family in murine macrophages --- p.87 / Chapter CHAPTER SIX --- ANTI-TUMOR ACTIVITIES ON MURINE TUMOR CELL LINES OF KLEBSIELLA K1 &K24 CAPSULAR POLYSACCHARIDES --- p.94 / Chapter 6.1. --- Effect of K1 and K24 Capsular Polysaccharides on the In vivo Anti-tumor Activities on EAT-bearing Mice --- p.94 / Chapter 6.2. --- In vivo Anti-tumor Effect on the Survival of EAT-Bearing Mice by Treatment of K1 and K24 CPS --- p.97 / Chapter CHAPTER SEVEN --- "ANTI-TUMOR ACTIVITIES OF COMBINED TREATMENT OF KLEBSIELLA K1 &K24 CAPSULAR POLYSACCHARIDES AND CHEMOTHERAPEUTIC DRUGS: ACTINOMYCIN D, CYCLOPHOSPHAMIDE AND DOXORUBICIN" --- p.100 / Chapter 7.1. --- "Cytotoxic Effect Chemotherapeutic Drugs： Actinomycin D (Act D), Cyclophosphamide (CY) and Doxorubicin (Dox), on WEHI-164 cell line" --- p.100 / Chapter 7.2. --- "Cytotoxic Effect of the Combined Treatment of the CPS and the Chemotherapeutic Drugs (Act D, CY and Dox), on WEHI-164 cell line" --- p.104 / Chapter 7.3. --- "Anti-Tumor Effect of Chemotherapeutic Drugs (Act D, CY and Dox) on EAT-bearing Mice in vivo" --- p.111 / Chapter 7.4. --- "Combined Treatment of the CPS and Chemotherapeutic Drugs (Act D, CY and Dox) on the EAT Growth in vivo" --- p.115 / Chapter 7.5. --- "Combined Treatment of the CPS and Chemotherapeutic Drugs (Act D, CY and Dox) on the Survival of EAT-bearing Mice in vivo" --- p.122 / Chapter CHAPTER EIGHT --- GENERAL DISCUSSION --- p.135 / Chapter 8.1. --- "Extraction, Purification and Characterization of K1 and K24 Capsular Polysaccharides from Klebsiella pneumoniae Serotype 1 and Serotype 24" --- p.135 / Chapter 8.2. --- Effect of K1 and K24 Capsular Polysaccharides on the Macrophage Activation --- p.140 / Chapter 8.3. --- Anti-tumor Activities on Murine Tumor Cell LInes of Klebsiella K1 and K24 Capsular Polysaccharides --- p.145 / Chapter 8.4. --- "Anti-tumor Activities of Combined Treatment of K1 and K24 Capsular Polysaccharides and Chemotherapeutic Drugs： Actinomycin D, Cyclophosphamide and Doxorubicin" --- p.148 / Chapter CHAPTER NINE --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.152 / BIBLIOGRAPHY --- p.156

Choi Pui-yu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 113-123). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese Version) --- p.iii / Acknowledgements --- p.v / List of Abbreviations --- p.vi / Table of Contents --- p.viii / List of Tables --- p.xii / List of Figures --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Literature Review / Chapter 2.1 --- Mushroom Polysaccharides with Antitumor Activities --- p.7 / Chapter 2.1.1 --- Antitumor β-Glucans --- p.7 / Chapter 2.1.2 --- Antitumor Heteroglycans --- p.9 / Chapter 2.1.3 --- Antitumor Polysaccharide-Protein Complexes --- p.12 / Chapter 2.2 --- Antitumor Activities and Structural Characteristics of Mushroom Polysaccharides --- p.15 / Chapter 2.3 --- Antitumor Effects of Mushroom Polysaccharides In Vitro --- p.19 / Chapter 2.4 --- Antitumor Effects of Mushroom Polysaccharides In Vivo --- p.21 / Chapter 2.5 --- Immunomodulatory Activities --- p.24 / Chapter 2.6 --- Activation of Cytokines by Mushroom Polysaccharides --- p.28 / Chapter 2.7 --- Induction of Nitric Oxide Synthase by Mushroom Polysaccharides --- p.32 / Chapter 2.8 --- Tremella aurantialba --- p.34 / Chapter Chapter 3 --- Materials and Methods --- p.35 / Chapter 3.1 --- Extraction --- p.35 / Chapter 3.1.1 --- Extraction of Crude Tremella aurantialba Polysaccharide --- p.35 / Chapter 3.1.2 --- Fractionation --- p.38 / Chapter 3.1.3 --- Polysaccharide and Protein Content Determination --- p.38 / Chapter 3.1.3.1 --- Phenol-Sulfuric Assay --- p.39 / Chapter 3.1.3.2 --- Lowry-Folin Method --- p.39 / Chapter 3.1.4 --- Gas Chromatography (GC) --- p.40 / Chapter 3.1.5 --- Modified Carbazole Assay --- p.41 / Chapter 3.1.6 --- High Performance Liquid Chromatography (HPLC) --- p.42 / Chapter 3.2 --- In Vitro Studies --- p.43 / Chapter 3.2.1 --- Maintenance of Cell Lines --- p.43 / Chapter 3.2.2 --- Effect on Cancer Cell Lines --- p.43 / Chapter 3.2.2.1 --- Trypan Blue Exclusion Methods --- p.44 / Chapter 3.2.2.2 --- MTT Assay --- p.44 / Chapter 3.2.3 --- Effect on Normal Cell Line --- p.45 / Chapter 3.2.4 --- Coulter Counter --- p.46 / Chapter 3.3 --- In Vivo Studies --- p.47 / Chapter 3.3.1 --- Animals --- p.47 / Chapter 3.3.2 --- Maintenance of Sarcoma 180 Cell Line --- p.47 / Chapter 3.3.3 --- Effect of TAP Fractions on Sarcoma 18 Solid Tumor --- p.48 / Chapter 3.3.3.1 --- Injection of TAP Fractions 24 h After Transplantation --- p.48 / Chapter 3.3.3.2 --- Injection of TAP Fractions 72 h After Transplantation --- p.49 / Chapter 3.4 --- Effect of TAP Fractions on Modulating mRNA Expression of Cytokines and Nitric Oxide Synthase --- p.51 / Chapter 3.4.1 --- Treatment of Mice --- p.51 / Chapter 3.4.2 --- Isolation of Splenocytes and Peritoneal Exduate Cells --- p.51 / Chapter 3.4.3 --- Extraction of Total mRNA from Splenocyte and Peritoneal Exduate Cells --- p.52 / Chapter 3.4.4 --- Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.53 / Chapter 3.4.4.1 --- Reverse Transcription --- p.53 / Chapter 3.4.4.2 --- Polymerase Chain Reaction --- p.56 / Chapter 3.4.5 --- DNA Sequencing --- p.57 / Chapter 3.5 --- Statistical Analysis --- p.58 / Chapter Chapter 4 --- Results --- p.59 / Chapter 4.1 --- Isolation and Characterization of TAP Fractions --- p.59 / Chapter 4.1.1 --- Percentage Yield of TAP Fractions --- p.59 / Chapter 4.1.2 --- Polysaccharide and Protein Content of TAP Fractions --- p.59 / Chapter 4.1.3 --- Relative Monosaccharide Contents in TAP Fractions --- p.60 / Chapter 4.1.4 --- Results of HPLC --- p.60 / Chapter 4.2 --- Effects of TAP Fractions In Vitro --- p.69 / Chapter 4.2.1 --- Effects of TAP Fractions on Suspension Cancer Cell Lines --- p.69 / Chapter 4.2.2 --- Effects of TAP Fractions on Adhesion Cancer Cell Lines --- p.69 / Chapter 4.2.3 --- Effects of TAP Fractions on Normal Cell Line --- p.70 / Chapter 4.2.4 --- Effect of TAP 2 on HL-60 Cell Line as Evaluated by Coulter Counter --- p.70 / Chapter 4.3 --- Antitumor Effect of TAP Fractions In Vivo --- p.78 / Chapter 4.4 --- Effect of TAP Fractions on Modulating mRNA Expressions of Cytokines and Nitric Oxide Sythase (NOS) --- p.83 / Chapter 4.4.1 --- Results of RT-PCR --- p.83 / Chapter 4.4.2 --- Sequencing --- p.84 / Chapter Chapter 5 --- Discussion --- p.91 / Chapter 5.1 --- Characterization of TAP Fractions --- p.91 / Chapter 5.2 --- Antitumor Effects of TAP Fractions In Vitro --- p.96 / Chapter 5.3 --- Furhter Study --- p.109 / Chapter Chapter 6 --- Conclusion --- p.111 / References --- p.113

Poon Shuk-ching. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 113-125). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract (Chinese Version) --- p.iv / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xiii / Chapter Chapterl 1 --- ntroduction --- p.1 / Chapter Chapter 2 --- Literature Review --- p.5 / Chapter 2.1 --- Mushroom Polysaccharides From Basidiomycetes --- p.5 / Chapter 2.1.1 --- Antitumor and Immunomodulatory Activity --- p.6 / Chapter 2.1.2 --- Antiviral Activity --- p.9 / Chapter 2.1.3 --- Hypoglycermic Activity --- p.11 / Chapter 2.1.4 --- Free Radical Scavenging Activity --- p.11 / Chapter 2.2 --- Mushroom Dictyophora indusiata --- p.13 / Chapter 2.2.1 --- Nutritional Value --- p.13 / Chapter 2.2.2 --- Structural Characteristic of Dictyophora indusiata Polysaccharides --- p.14 / Chapter 2.2.3 --- Biological Activity --- p.16 / Chapter 2.3 --- In vivo Antitumor Study --- p.19 / Chapter 2.4 --- Induction of Cytokines Production in Immune System --- p.21 / Chapter 2.5 --- In vitro Antitumor Study --- p.25 / Chapter 2.6 --- Cell Cycle Regulation --- p.28 / Chapter Chapter 3 --- Materials & Methods --- p.34 / Chapter 3.1 --- Extraction --- p.34 / Chapter 3.1.1 --- Extraction of Dictyophora indusiata Polysaccharides --- p.34 / Chapter 3.1.2 --- Purification of Dictyophora indusiata Polysaccharides --- p.35 / Chapter 3.1.2.1 --- Preparation of DEAE-cellulose Ion Exchanger --- p.35 / Chapter 3.1.2.2 --- Fractionation --- p.35 / Chapter 3.2. --- Characterization of Dictyophora indusiata Polysaccharides --- p.39 / Chapter 3.2.1 --- Polysaccharide Content Determination --- p.39 / Chapter 3.2.2 --- Protein Content Determination --- p.39 / Chapter 3.2.3 --- Gas Chromatography (GC) --- p.40 / Chapter 3.2.4 --- Uronic Acid Content Determination --- p.42 / Chapter 3.2.5 --- High Performance Liquid Chromatography (HPLC) --- p.43 / Chapter 3.3 --- In vivo Studies --- p.44 / Chapter 3.3.1 --- Animals --- p.44 / Chapter 3.3.2 --- Maintenance of Sarcoma 180 Cell Line --- p.44 / Chapter 3.3.3 --- Effect of DI3 Fraction on Sarcoma 180 Solid Tumor --- p.45 / Chapter 3.3.4 --- Effect of DI3c Fraction on Tumor Necrosis Factor-Alpha (TNF-α) and Interleukin 2 (IL-2) Production --- p.47 / Chapter 3.3.4.1 --- Treatment of Mice --- p.47 / Chapter 3.3.4.2 --- Preparation of Mouse Serum --- p.47 / Chapter 3.3.4.3 --- Enzyme-linked Immunosorbent Assay (ELISA) for TNF-α Production --- p.48 / Chapter 3.3.4.4 --- Enzyme-linked Immunosorbent Assay (ELISA) for IL-2 Production --- p.49 / Chapter 3.4 --- In vitro Studies --- p.51 / Chapter 3.4.1 --- Maintenance of Cell Lines --- p.51 / Chapter 3.4.2 --- Effect on Cancer Cell Lines --- p.52 / Chapter 3.4.3 --- Cytotoxicity on Normal Cell Line --- p.52 / Chapter 3.4.4 --- Trypan Blue Exclusion Method --- p.53 / Chapter 3.4.5 --- MTT Assay --- p.54 / Chapter 3.4.6 --- BrdU Incorporation --- p.55 / Chapter 3.5 --- Statistical Analysis --- p.56 / Chapter Chapter 4 --- Results --- p.57 / Chapter 4.1 --- Extraction and Fractionation of Polysaccharides from Dictyophora indusiata --- p.57 / Chapter 4.1.1 --- Percentage Yield of Crude DI Polysaccharides --- p.57 / Chapter 4.1.2 --- Percentage Yield of DI3 Fractions --- p.57 / Chapter 4.2 --- Characterization of DI3 Fractions --- p.62 / Chapter 4.2.1 --- Polysaccharide and Protein Contents of DI3 Fractions --- p.62 / Chapter 4.2.2 --- Relative Monosaccharide and Uronic Acid Content in Different DI3 Fractions --- p.62 / Chapter 4.2.3 --- Estimated Molecular Weight of DI3 Fractions --- p.65 / Chapter 4.3 --- Antitumor Effect of Dictyophora indusiata Polysaccharides In vivo --- p.70 / Chapter 4.3.1 --- In vivo Antitumor Effect of Crude DI Polysaccharides --- p.70 / Chapter 4.3.2 --- In vivo Antitumor Effect of Various Fractions of DI3 --- p.70 / Chapter 4.3.3 --- Effect of DI3c on TNP-α and IL-2 Production in Mice --- p.78 / Chapter 4.4 --- In vitro Effects of DI3 Fractions on Cell Density and Viability on Normal and Cancer Cell Lines --- p.86 / Chapter 4.4.1 --- Effects of DI3 Fractions on Cell Density and Viability of Human Leukemic HL-60 and K-562 and Mouse Sarcoma 180 Cells --- p.86 / Chapter 4.4.2 --- Effects of DI3 Fractions on the Growth of Human Liver Cancer HepG2 and Normal Monkey Kidney Vero Cells --- p.86 / Chapter 4.4.3 --- Effect of DI3b Fraction on Proliferation of HL-60 Cells Determined by BrdU Incorporation --- p.94 / Chapter Chapter 5 --- Discussions --- p.96 / Chapter 5.1 --- Extraction and Characterization of DI3 Fractions --- p.96 / Chapter 5.2 --- Antitumor Effects of Dictyophora indusiata Polysaccharides --- p.101 / Chapter 5.3 --- Further Studies --- p.109 / Chapter Chapter 6 --- Conclusion --- p.111 / References --- p.113