by Fu Sai-chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 102-111). / Acknowledgements / List of Abbreviation / Abstract / Prologue --- p.1 / Chapter / Chapter 1. --- Introduction --- p.2 / Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4 / Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6 / Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6 / Chapter 1.3.1 --- The triterpenoids --- p.7 / Chapter 1.3.2 --- Matrix polysaccharides --- p.8 / Chapter 1.4 --- Medicinal properties and biological activities --- p.9 / Chapter 1.4.1 --- Medicinal properties --- p.9 / Chapter 1.4.2 --- Biological activities --- p.10 / Chapter 1.4.2.1 --- Anti-tumour activity --- p.10 / Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11 / Chapter 1.4.2.3 --- Effects on nervous system --- p.11 / Chapter 1.4.2.4 --- Effects on respiratory system --- p.12 / Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12 / Chapter 1.4.2.6 --- Effects on immune system --- p.14 / Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14 / Chapter 1.4.3 --- Summary --- p.15 / Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15 / Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Cell culture reagents / Chapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20 / Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20 / Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20 / Chapter 2.1.4 --- Mitogens and test samples --- p.21 / Chapter 2.1.5 --- Trypan blue exclusion test --- p.21 / Chapter 2.1.6 --- Liquid scintillation counting --- p.21 / Chapter 2.2 --- Animal care --- p.22 / Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22 / Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23 / Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23 / Chapter 2.4.2 --- Preparation of liver homogenates --- p.23 / Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25 / Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25 / Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25 / Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26 / Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26 / Chapter 2.5 --- Preparation of murine hepatocytes --- p.27 / Chapter 2.5.1 --- Liver perfusion --- p.27 / Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29 / Chapter 2.5.3 --- Incubation protocol --- p.29 / Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30 / Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31 / Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31 / Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31 / Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32 / Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34 / Chapter 2.7 --- Chromatographic methods --- p.34 / Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34 / Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35 / Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35 / Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36 / Chapter 2.8 --- Measurement of in vivo antibody production --- p.39 / Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39 / Chapter 2.8.2 --- Direct plaque assay --- p.39 / Chapter 2.9 --- Biochemical analysis --- p.39 / Chapter 2.9.1 --- Determination of protein content --- p.40 / Chapter 2.9.2 --- Determination of hexose content --- p.40 / Chapter 2.9.3 --- Determination of uronic acid content --- p.40 / Chapter 2.9.4 --- Determination of sulphate content --- p.41 / Chapter 2.9.5 --- Determination of hexosamine content --- p.42 / Chapter 2.10 --- Statistical analysis --- p.42 / Chapter 3. --- Results --- p.44 / Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44 / Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47 / Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47 / Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47 / Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51 / Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51 / Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51 / Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55 / Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55 / Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55 / Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58 / Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65 / Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65 / Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69 / Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69 / Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72 / Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76 / Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76 / Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76 / Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80 / Chapter 4. --- Discussions --- p.83 / Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83 / Chapter 4.2 --- The significance of enhancement of GSH status --- p.89 / Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95 / Conclusions --- p.99 / Appendix --- p.100 / Reference --- p.102

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 Chen Paul. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 141-150). / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1 --- Immunomodulation --- p.1 / Chapter 1.2 --- Effector cells mediating anti-tumour immunity --- p.1 / 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 (LAK) --- p.6 / Chapter 1.3 --- Cytokines as immunomodulators in cancer therapy --- p.7 / Chapter 1.3.1 --- Tumour Necrosis Factor-α (TNF-α) --- p.7 / Chapter 1.3.2 --- Interleukin-1 (IL-1) --- p.9 / Chapter 1.3.3 --- Interleukin-2 (IL-2) --- p.9 / Chapter 1.3.4 --- Granulocytes/Macrophages Colony-Stimulating Factors --- p.10 / Chapter 1.4 --- Polysaccharides as potential immunostimulating agents --- p.11 / Chapter 1.5 --- General properties of Klebsiella pneumoniae --- p.12 / Chapter 2. --- AIM AND SCOPE OF THIS DISSERTATION --- p.16 / Chapter 3. --- MATERIALS AND METHODS --- p.18 / Chapter 3.1 --- Materials --- p.18 / Chapter 3.1.1 --- Animals --- p.18 / Chapter 3.1.2 --- Klebsiella pneumoniae K24 --- p.18 / Chapter 3.1.3 --- Cell lines --- p.18 / Chapter 3.1.4 --- "Buffer, Culture media and Chemicals" --- p.19 / Chapter 3.2 --- Methods --- p.27 / Chapter 3.2.1 --- Extraction and Characterization of Klebsiella pneumoniae K24 Capsular Polysaccharide (K24 CPS) --- p.27 / Chapter 3.2.2 --- Assays of Immunomodulatory Activities of K24 CPS on Lymphocytes --- p.30 / Chapter 3.2.3 --- Assays of Immunomodulatory Effect of K24 CPS on Macrophages --- p.34 / Chapter 3.2.4 --- Assays of Anti-Tumour Activities of K24 CPS --- p.39 / Chapter 3.2.5 --- Assays of the Effects of K24 CPS on the Proliferation and Differentiation of Murine Bone Marrow Cells --- p.54 / Chapter 3.2.6 --- Assays of the Immunorestorative Activities of K24 CPS --- p.56 / Chapter 4. --- EXTRACTION AND CHARACTERIZATION OF KLEBSIELLA PNEUMONIAE K24 CAPSULAR POLYSACCHARIDE (K24 CPS) --- p.59 / Chapter 4.1 --- Preparation of Klebsiella pneumoniae K24 CPS Capsular Polysaccharide (K24 CPS) --- p.59 / Chapter 4.2 --- Acetic Acid Treatment of K24 CPS --- p.59 / Chapter 4.3 --- Gel Filtration --- p.59 / Chapter 4.4 --- Carbohydrate and Protein contents of K24 CPS --- p.61 / Chapter 4.5 --- Cytotoxicity Assay using Artemia franciscana (Brine Shrimp) --- p.61 / Chapter 5. --- IMMUNOMODULATORY EFFECTS OF K24 CPS --- p.68 / Chapter 5.1 --- The Effect of K24 CPS in vitro Mitogenic Assay of K24 CPS using Murine Splenocytes --- p.68 / Chapter 5.2 --- The in vivo Mitogenic Effect of K24 CPS on Murine Splenic Lymphocytes --- p.73 / Chapter 5.3 --- The Effect of K24 CPS on the Production of Interleukin-2 (IL-2)-like substance by Murine Splenocytes --- p.73 / Chapter 5.4 --- The effect of K24 CPS on the in vitro Stimulation of Murine Macrophage Nitric Oxide (NO) Production --- p.73 / Chapter 5.5 --- The effect of K24 CPS on the in vitro Stimulation of Macrophage Interleukin-1-like Production --- p.77 / Chapter 5.6 --- The effect of K24 CPS on in vivo Migration of Macrophage --- p.82 / Chapter 5.7 --- The effect of K24 CPS in vitro Stimulation of Macrophage Tumour Necrosis Factor- a (TNF-a) Production --- p.82 / Chapter 6. --- IN VITRO ANTI-TUMOUR EFFECT OF K24 CPS --- p.89 / Chapter 6.1 --- The in vitro Cytostatic effect of K24 CPS on the Suppression of EAT growth --- p.89 / Chapter 6.2 --- The effect of K24 CPS on cell cycle of EAT cells --- p.89 / Chapter 6.3 --- Study of the cytostatic effect of K24 CPS on EAT cells using Western Analysis --- p.93 / Chapter 6.3.1 --- Pattern of Phosphotyrosine Proteins --- p.93 / Chapter 6.3.2 --- Pattern of Phosphoserine Proteins --- p.96 / Chapter 6.3.3 --- Pattern of Phosphothreonine Proteins --- p.96 / Chapter 6.3.4 --- Level of c-fos --- p.99 / Chapter 6.3.5 --- Level of c-jun --- p.99 / Chapter 6.3.6 --- Level of c-myc --- p.102 / Chapter 7. --- THE IN VIVO ANTI-TUMOUR ACTIVITIES OF K24 CPS --- p.103 / Chapter 7.1 --- The effect of K24 CPS on the In vivo Suppression of EAT growth --- p.103 / Chapter 7.2 --- The effect of K24 CPS on the survival of EAT-bearing mice --- p.103 / Chapter 7.3 --- The effect of K24 CPS on the in vivo induction of Natural Killer (NK) Cell Cytotoxicity --- p.111 / Chapter 7.4 --- The effect of K24 CPS in vitro induction of Lymphokine-activated Killer (LAK) Cell Cytotoxicity --- p.111 / Chapter 7.5 --- The effect of K24 CPS on the in vivo Induction of Lymphokine-activated Killer (LAK) Cell Cytotoxicity --- p.114 / Chapter 7.6 --- The effect of K24 CPS on the endogenous production of TNF-α --- p.114 / Chapter 7.7 --- The effect of K24 CPS on the endogenous TNF-α production and EAT growthin vivo --- p.117 / Chapter 8. --- THE IMMUNORESTORATIVE ACTIVITIES OF K24 CPS --- p.122 / Chapter 8.1 --- The in vivo Immunorestorative Activities of K24 CPS in EAT-bearing Mice --- p.122 / Chapter 8.2 --- The in vitro Immunorestorative Activities of K24 CPS in Mice bearing 10-day-old- EAT --- p.122 / Chapter 9. --- THE EFFECT OF K24 CPS IN VITRO INDUCTION OF MURINE BONE MARROW CELLS PROLIFERATION AND DIFFERENTIATION --- p.126 / Chapter 9.1 --- The effect of K24 CPS in vitro induction of Murine Bone Marrow Cells Proliferation --- p.126 / Chapter 9.2 --- The effect of K24 CPS in vitro induction of Murine Bone Marrow Cells Differentiation --- p.126 / Chapter 10. --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.135 / Chapter 11. --- BIBLIOGRAPHY --- p.141

by Kok Dick Shun, Louis. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references. / Acknowledgements --- p.I / Table of Contents --- p.II / Abbreviations --- p.V / Aim and Scope of This Dissertation --- p.IX / Abstract --- p.X / Chapter Chapter One： --- General Introduction --- p.1 / Chapter 1.1 --- An Overview of the Immune System --- p.2 / Chapter 1.1.1 --- Innate Immunity --- p.2 / Chapter 1.1.2 --- Adaptive Immunity --- p.3 / Chapter 1.1.2.1 --- Humoral antibody immune response --- p.4 / Chapter 1.1.2.2 --- Cell- mediated immune response --- p.5 / Chapter 1.2 --- Immunomodulation --- p.6 / Chapter 1.3 --- An overview of the Host-mediated response against tumours --- p.9 / Chapter 1.3.1 --- T and B lymphocytes --- p.9 / Chapter 1.3.2 --- M acrophages --- p.14 / Chapter 1.3.3 --- Natural killer cells --- p.17 / Chapter 1.3.4 --- Lymphokines-activated killer cells --- p.20 / Chapter 1.3.5 --- Tumour infiltrating lymphocytes --- p.22 / Chapter 1.3.6 --- Cytokines --- p.23 / Chapter 1.4 --- Carbohydrates as Potential Immunostimulating agents --- p.33 / Chapter 1.5 --- General Properties of Bupleuri radix (B.R.) --- p.35 / Chapter Chapter Two： --- Materials and Methods --- p.36 / Chapter 2.1 --- Materials --- p.37 / Chapter 2.1.1 --- Animals --- p.37 / Chapter 2.1.2 --- Bupleuri radix --- p.37 / Chapter 2.1.3 --- "Buffers, culture media and chemicals" --- p.37 / Chapter 2.1.4 --- Cell lines --- p.48 / Chapter 2.2 --- Methods --- p.49 / Chapter 2.2.1 --- Extraction and fractionation of Bupleuri radix --- p.49 / Chapter 2.2.2 --- Purification of Bupleuri radix --- p.54 / Chapter 2.2.3 --- Characterization of Bupleuri radix --- p.55 / Chapter 2.2.4 --- In vivo Drug Treatment --- p.59 / Chapter 2.2.5 --- Isolation and preparation of cells --- p.59 / Chapter 2.2.6 --- Assays for the immunomodulatory activities of Bupleuri radix --- p.62 / Chapter 2.2.7 --- Assays for the immunorestorative properties of Bupleuri radix --- p.74 / Chapter 2.2.8 --- Assays for the anti-tumour activities of Bupleuri radix --- p.75 / Chapter 2.2.9 --- Statistical analysis --- p.83 / Chapter Chapter Three： --- "Fractionation, Purification and Characterization of Bioactive Compounds from Bupleuri radix" --- p.84 / Chapter 3.1 --- Results / Chapter 3.1.1 --- Extraction and Fractionation of Bupleuri radix --- p.85 / Chapter 3.1.2 --- Purification of Bupleuri radix --- p.85 / Chapter 3.1.3 --- Carbohydrate and Protein Contents of B.R. Fractions --- p.87 / Chapter 3.1.4 --- Lack of cytotoxicity of Bupleuri radix to Mouse Splenocytes --- p.91 / Chapter 3.1.5 --- LC50 of B.R. Fractions determined by Brine Shrimp Bioassay --- p.91 / Chapter 3.1.6 --- Heat stability of B.R. Fractions --- p.93 / Chapter 3.1.7 --- "Uronic Acid Content of BRIai, BRIaii, BRIbi and BRIbii" --- p.93 / Chapter 3.2 --- Discussion --- p.93 / Chapter Chapter Four： --- The Immunomodulatory Activities of Bupleuri radix --- p.96 / Chapter 4.1 --- Results / Chapter 4.1.1 --- Effect of Bupleuri radix on the Specific and Nonspecific Immunity --- p.97 / Chapter 4.1.1.1 --- Mitogenic effect of B.R. Fractions on Murine Splenocytes in vitro --- p.97 / Chapter 4.1.1.2 --- Mitogenic effect of B.R. Fractions on Murine Splenocytes ex vivo --- p.97 / Chapter 4.1.1.3 --- In vitro Mitogenic effect of B.R. Fractions treated with Periodate --- p.103 / Chapter 4.1.1.4 --- In vitro Mitogenic effect of B.R. Fractions treated with Acetic Acid --- p.103 / Chapter 4.1.1.5 --- In vitro Co -mitogenic effect of B.R. Fractions with Polymyxin B Sulphate --- p.107 / Chapter 4.1.1.6 --- Effect of B.R. Fractions on Lymphocyte sub-populations --- p.107 / Chapter 4.1.1.7 --- Primary Humoral Immune Response to SRBC in B.R.-treated mice --- p.107 / Chapter 4.1.1.8 --- Activity of cytotoxic T cells in B.R-treated mice --- p.111 / Chapter 4.1.1.9 --- Effect of B.R. Fractions on Interleukin-1 - like Factors Production --- p.111 / Chapter 4.1.1.10 --- Effect of B.R. Fractions on Interleukin-2 Production --- p.116 / Chapter 4.1.1.11 --- Effect of B.R. Fractions on Interleukin-2 Receptor Expression on Murine Splenocytes --- p.116 / Chapter 4.1.1.12 --- Effect of B.R. Fractions on GM-CSF Production --- p.119 / Chapter 4.1.1.13 --- Immunopotentiating effects of B.R. Fractions on Macrophages: --- p.119 / Chapter 4.1.1.13.1 --- In vivo Migration of Macrophages in B.R.-treated mice --- p.119 / Chapter 4.1.1.13.2 --- Effect of B.R. Fractions on the Fc Receptor Expression on Murine Resident Peritoneal Exudate Cells --- p.123 / Chapter 4.1.2 --- Immunorestorative Properties of Bupleuri radix --- p.123 / Chapter 4.1.2.1 --- Effect of B.R. Fractions on Lymphocyte Blastogenesis in Aged Mice --- p.123 / Chapter 4.1.2.2 --- Effect of B.R. Fractions on Lymphocyte Blastogenesis in Tumour-bearing Mice --- p.125 / Chapter 4.2 --- Discussion --- p.125 / Chapter Chapter Five： --- The Anti-tumour Activities of Bupleuri radix --- p.132 / Chapter 5.1 --- Results / Chapter 5.1.1 --- Cytostatic Effect of B.R. Fractions on Murine Tumour Cell Lines in vitro --- p.133 / Chapter 5.1.2 --- Effect of B.R. Fractions on the Growth of Tumour Ceils in vivo --- p.133 / Chapter 5.1.3 --- Effect of B.R. Fractions on the Survival of EAT-bearing mice --- p.140 / Chapter 5.1.4 --- Ex vivo Induction of Natural Killer Cell Activity by B.R. Fractions --- p.146 / Chapter 5.1.5 --- In vitro Induction of Lymphokine-activated Killer Cell Activity by B.R Fractions --- p.149 / Chapter 5.1.6 --- In vivo Induction of Tumour Infiltrating Lymphocytes by B.R. Fractions --- p.149 / Chapter 5.1.7 --- In vitro Induction of Macrophage-mediated Cytostatic Effect on Tumour Cells by B.R. Fractions --- p.151 / Chapter 5.1.8 --- In vitro Induction of Macrophage-mediated Cytostatic Eifect on Tumour Cells by B.R. Fractions --- p.153 / Chapter 5.1.9 --- Effect of B.R. Fractions on γ-interferon Production in vitro --- p.156 / Chapter 5.2 --- Discussion --- p.156 / Chapter Chapter Six： --- "General Discussion, Conclusion and Future Prospects" --- p.164 / Bibliography --- p.i