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Immunomodulatory and anti-tumor activities of K1 capsular polysaccharide from klebsiella pneumoniae.

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

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_321868
Date January 1997
ContributorsHo, Cheong Yip., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, vii, 164 leaves : ill. ; 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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