Tang Chi-Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 89-112). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vi / Table of contents --- p.viii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- History and prevalence of food allergy --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as a major shellfish allergen --- p.13 / Chapter 2.4 --- Use of animal model in the studies of food allergy --- p.22 / Chapter 2.5 --- Future approaches for treatment of food allergy --- p.27 / Chapter Chapter 3. --- Cloning and expression of recombinant tropomyosin --- p.30 / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Materials and Methods --- p.31 / Chapter 3.2.1 --- Design of PCR primers for amplification of tropomyosin gene --- p.31 / Chapter 3.2.2 --- Cloning of PCR-amplified cDNA into vector --- p.32 / Chapter 3.2.3 --- Transformation of competent E. coli Ml5 cells --- p.34 / Chapter 3.2.4 --- Confirmation of DNA sequence of the cloned vector --- p.34 / Chapter 3.2.5 --- Induction of the recombinant protein --- p.35 / Chapter 3.2.6 --- Purification and storage of the recombinant protein under native condition --- p.36 / Chapter 3.2.7 --- Concentration measurement and storage of the recombinant protein --- p.37 / Chapter 3.2.8 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.38 / Chapter 3.2.9 --- Regeneration of the Ni-NTA column --- p.40 / Chapter 3.3 --- Results and discussion --- p.42 / Chapter 3.3.1 --- DNA sequence of the cloned vector --- p.42 / Chapter 3.3.2 --- Expression of the recombinant protein --- p.42 / Chapter 3.3.3 --- Sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE) --- p.43 / Chapter Chapter 4. --- Induction of hypersensitive response to shrimp tropomyosin in mice --- p.47 / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Materials and methods --- p.52 / Chapter 4.2.1 --- Mice and reagents --- p.52 / Chapter 4.2.2 --- Animal sensitization and challenge --- p.53 / Chapter 4.2.3 --- Morphological and behavioral changes --- p.54 / Chapter 4.2.4 --- Tropomyosin-specific IgE level --- p.55 / Chapter 4.2.5 --- Passive cutaneous anaphylaxis (PCA) reaction --- p.56 / Chapter 4.2.6 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.56 / Chapter 4.2.7 --- Cytokine profiles of splenoctyes --- p.58 / Chapter 4.2.8 --- Histological examination of small intestine --- p.59 / Chapter 4.2.9 --- Statistical analysis --- p.59 / Chapter 4.3 --- Results --- p.63 / Chapter 4.3.1 --- Morphological and behavioral changes after challenge --- p.63 / Chapter 4.3.2 --- Tropomyosin-specific IgE level --- p.63 / Chapter 4.3.3 --- Passive cutaneous anaphylaxis (PCA) --- p.64 / Chapter 4.3.4 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.68 / Chapter 4.3.5 --- Cytokine profiles of splenocytes --- p.70 / Chapter 4.3.6 --- Histology of small intestines --- p.76 / Chapter 4.4 --- Discussion --- p.79 / Chapter Chapter 5. --- General conclusion --- p.88 / References --- p.89

Kung, Wing Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 92-115). / Abstracts in English and Chinese. / Abstract --- p.ii / Acknowledgements --- p.vii / Table of contents --- p.ix / List of Tables --- p.xii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- Food allergy and its prevalence --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as the major allergen in shellfish --- p.15 / Chapter 2.4 --- Cross reactivity and epitope mapping of tropomyosin --- p.21 / Chapter 2.5 --- Novel approaches for the treatment of food allergy --- p.29 / Chapter Chapter 3. --- Expression of shrimp recombinant tropomyosin and sensitization of mice --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Materials and Methods --- p.40 / Chapter 3.2.1 --- "Recovery of E, coli with tropomyosin-carrying plasmid" --- p.40 / Chapter 3.2.2 --- Preparation of tropomyosin-carrying plasmid --- p.41 / Chapter 3.2.3 --- Confirmation of DNA sequence of the tropomyosin --- p.41 / Chapter 3.2.4 --- Identification of the recombinant protein --- p.43 / Chapter 3.2.5 --- Purification of the recombinant protein --- p.43 / Chapter 3.2.6 --- Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.44 / Chapter 3.2.7 --- Concentration measurement of the recombinant tropomyosin --- p.45 / Chapter 3.2.8 --- Mice --- p.46 / Chapter 3.2.9 --- Mice sensitization and challenging --- p.46 / Chapter 3.2.10 --- Tropomyosin-specific IgE level in blood --- p.47 / Chapter 3.2.11 --- Statistical analysis --- p.49 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- DNA sequence of the cloned tropomyosin --- p.52 / Chapter 3.3.2 --- Expression and purification of tropomyosin --- p.52 / Chapter 3.3.3 --- Hypersensitivity symptoms after challenge --- p.53 / Chapter 3.3.4 --- Blood tropomyosin-specific IgE level --- p.53 / Chapter 3.4 --- Discussion --- p.62 / Chapter Chapter 4. --- Identification of T cell epitopes --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and methods --- p.67 / Chapter 4.2.1 --- Soluble epitope peptide synthesis --- p.68 / Chapter 4.2.2 --- Isolation of spleen cells from mice --- p.69 / Chapter 4.2.3 --- T cell proliferation assay --- p.70 / Chapter 4.3 --- Results --- p.71 / Chapter 4.3.1 --- Splenocyte proliferation to synthetic peptide --- p.72 / Chapter 4.3.2 --- Splenocyte proliferation to synthetic peptides pool --- p.72 / Chapter 4.4 --- Discussion --- p.77 / Chapter Chapter5 --- General conclusion --- p.89 / References --- p.92