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Immunomodulatory activities of mushroom sclerotial polysaccharides isolated from Polyporus rhinocerus mediated by antigen-presenting cells.

Choi, Man Wing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 126-139). / Abstracts in English and Chinese. / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Antigen presenting cells (APC) in Immune systems --- p.1 / Chapter 1.1.1 --- Dendritic cells --- p.2 / Chapter 1.1.1.1 --- Differentiation of dendritic cells in mice --- p.2 / Chapter 1.1.1.2 --- Maturation of dendritic cells --- p.3 / Chapter 1.1.1.3 --- Stimulation and polarization of T cells stimulated by dendritic cells --- p.6 / Chapter 1.1.2 --- Monocyte and macrophage --- p.7 / Chapter 1.1.2.1 --- Differentiation of monocyte and macrophage in humans --- p.7 / Chapter 1.1.2.2 --- Changes involved in differentiation of monocytes into macrophages --- p.9 / Chapter 1.2 --- "Isolation, structure and activity of mushroom polysaccharides" --- p.13 / Chapter 1.2.1 --- Sources of mushroom polysaccharides --- p.13 / Chapter 1.2.2 --- Extraction methods --- p.14 / Chapter 1.2.3 --- Structure-Activity Relationship (SAR) of mushroom polysaccharides --- p.15 / Chapter 1.2.4 --- Previous studies on immunomodulatory effects of mushroom sclerotial polysaccharides --- p.18 / Chapter 1.3 --- Recognition of β-glucan by specific receptors --- p.20 / Chapter 1.3.1 --- Complement Receptor 3 (CR3) --- p.22 / Chapter 1.3.1.1 --- Introduction of CR3 --- p.22 / Chapter 1.3.1.2 --- Expressions of CR3 to recognize fungi --- p.22 / Chapter 1.3.2 --- Dectin-1 --- p.24 / Chapter 1.3.2.1 --- Introduction of Dectin-1 --- p.24 / Chapter 1.3.2.2 --- Structure of Full-length Dectin-1 --- p.26 / Chapter 1.3.2.2.1 --- Isoforms of Dectin-1 in Mice --- p.28 / Chapter 1.3.2.2.2 --- Isoforms of Dectin-1 in Humans --- p.28 / Chapter 1.3.2.3 --- Immune responses triggered by of Dectin-1 --- p.29 / Chapter 1.3.3 --- Toll-like 2 receptor (TLR2) --- p.31 / Chapter 1.3.3.1 --- Introduction of TLR2 --- p.31 / Chapter 1.3.3.2 --- Structure of TLR2 --- p.33 / Chapter 1.3.3.3 --- Immune responses triggered by TLR2 --- p.34 / Chapter 1.4 --- Research Objectives --- p.35 / Chapter Chapter 2 --- Materials and Methods --- p.38 / Chapter 2.1 --- Materials --- p.38 / Chapter 2.1.1 --- Mushroom sclerotia --- p.38 / Chapter 2.1.1.1 --- Polysaccharide extraction from mushroom sclerotia --- p.38 / Chapter 2.1.2 --- Antibodies and reagents --- p.41 / Chapter 2.1.3 --- Human acute leukocyte monocytic cell line and culture medium --- p.42 / Chapter 2.1.4 --- Preparation of murine bone marrow-derived immature dendritic primary cells (immature BMDCs) --- p.43 / Chapter 2.2 --- Methods --- p.45 / Chapter 2.2.1 --- Chemical Analysis --- p.45 / Chapter 2.2.1.1 --- Measurement of monosaccharide profile --- p.45 / Chapter 2.2.1.1.1 --- Acid depolymerisation --- p.45 / Chapter 2.2.1.1.2 --- Neutral sugar derivatization --- p.45 / Chapter 2.2.1.1.3 --- Gas chromatography (GC) --- p.46 / Chapter 2.2.1.2 --- Determination of total sugar by phenol-sulfuric acid method --- p.47 / Chapter 2.2.1.3 --- Determination of protein content by Lowry-Folin Method --- p.48 / Chapter 2.2.1.4 --- Size exclusion chromatography by high pressure liquid chromatography (HPLC) --- p.49 / Chapter 2.2.1.5 --- Endotoxin detection --- p.50 / Chapter 2.2.2 --- Measurement of Bioactivities --- p.51 / Chapter 2.2.2.1 --- Trypan blue exclusion assay --- p.51 / Chapter 2.2.2.2 --- MTT cell proliferation assay --- p.51 / Chapter 2.2.2.3 --- BrdU cell proliferation assay --- p.53 / Chapter 2.2.2.4 --- Expression of cell surface markers --- p.54 / Chapter 2.2.2.5 --- Phagocytosis / Endocytosis of FITC-labeled dextrans --- p.55 / Chapter 2.2.2.6 --- Nitric oxide production assay --- p.55 / Chapter 2.2.2.7 --- Reactive oxygen species production --- p.57 / Chapter 2.2.2.8 --- Determination of cytokine profile using cytokine antibody array --- p.58 / Chapter 2.2.2.9 --- Cell cycle analysis --- p.59 / Chapter 2.2.2.10 --- Expression of surface receptors --- p.60 / Chapter 2.2.2.11 --- Statistical analysis --- p.61 / Chapter Chapter 3 --- Results and Discussion --- p.61 / Chapter 3.1 --- Chemical characteristics of sclerotial polysaccharides --- p.61 / Chapter 3.1.1. --- The yield of sclerotial polysaccharides --- p.61 / Chapter 3.1.2 --- Total carbohydrate content of sclerotial polysaccharides --- p.65 / Chapter 3.1.3 --- Protein content of sclerotial polysaccharides --- p.66 / Chapter 3.1.4 --- Monosaccharide profiles of sclerotial polysaccharides from PR by gas chromatography (GC) --- p.66 / Chapter 3.1.5 --- Molecular weight of sclerotial polysaccharides from PR by size exclusion chromatography (SEC) --- p.69 / Chapter 3.1.6 --- Endotoxin test --- p.73 / Chapter 3.2 --- Immune responses for human monocytic cell line THP-1 --- p.74 / Chapter 3.2.1 --- MTT cell viability assay --- p.74 / Chapter 3.2.2 --- BrdU cell proliferation assay --- p.75 / Chapter 3.2.3 --- Change in cell morphology of THP-1 --- p.79 / Chapter 3.2.4 --- Phenotypic maturation of THP-1 --- p.81 / Chapter 3.2.5 --- Up-regulated phagocytic ability of THP-1 --- p.84 / Chapter 3.2.6 --- Increased nitrite production in THP-1 --- p.86 / Chapter 3.2.7 --- Production of reactive oxygen species --- p.88 / Chapter 3.2.8 --- Human cytokines profile array --- p.90 / Chapter 3.2.9 --- Cell cycle analysis --- p.93 / Chapter 3.2.10 --- Surface receptors expression --- p.95 / Chapter 3.2.11 --- Summary --- p.98 / Chapter 3.3 --- Immune responses for murine immature BMDCs --- p.102 / Chapter 3.3.1 --- Inhibition effects on murine immature BMDCs --- p.102 / Chapter 3.3.2 --- Change in cell morphology of murine immature BMDCs --- p.103 / Chapter 3.3.3 --- Phenotypic maturation of murine immature BMDCs --- p.105 / Chapter 3.3.4 --- Down-regulation of endocytosis in murine immature BMDCs --- p.106 / Chapter 3.3.5 --- Increased nitrite production --- p.109 / Chapter 3.3.6 --- Decreased expression of CD 11c in PRW-treated immature BMDCs --- p.109 / Chapter 3.3.7 --- Cytokine profile detection --- p.112 / Chapter 3.3.8 --- Surface receptors expression --- p.116 / Chapter 3.3.9 --- Summary --- p.119 / Chapter Chapter 4 --- Conclusion and future works --- p.123 / Appendix --- p.125 / References --- p.126

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_327133
Date January 2010
ContributorsChoi, Man Wing., Chinese University of Hong Kong Graduate School. Division of Life Sciences.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xx, 139 leaves : ill. (chiefly col.) ; 30 cm.
RightsUse 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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