This thesis examines the use of an antibody and single chain antibody to target myofibroblasts, for the diagnosis and treatment of liver fibrosis. Furthermore the effects of myofibroblasts depletion on inflammation, fibrosis and regeneration are investigated in animal models of chronic and acute liver disease. Liver myofibroblasts have become a focus as a potential therapeutic target as removal or prevention of their activation attenuates liver fibrosis severity. One of the major issues surrounding fibrosis treatment is developing a therapeutic that targets myofibroblasts without causing additional damage to the liver. Using an antibody may provide a solution due to their high specificity for their target antigen. Several antibodies are currently used in the clinic to treat a range of diseases. However no treatment is currently indicated for fibrosis, with transplantation being the only option at the later stages of disease. Our lab has developed a single chain antibody fragment (scab) capable of targeting activated myofibroblasts. Here we examine the use of this antibody as a drug delivery method <i>in vitro</i> and <i>in vivo</i>. This thesis also describes the generation of a human monoclonal IgG to target myofibroblasts and recruit the natural immune effector functions as a safer alternative to toxic drug conjugates. The presence of myofibroblasts is indicative for fibrosis and as such we have used myofibroblasts as a potential imaging target. Using C1-3 conjugated to a fluorophore, we investigated the possibility of grading fibrosis severity, using histological and imaging techniques.

Lee, Ngan Hon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 120-133). / Abstracts in English and Chinese. / Acknowledgements --- p.IV / Abstract --- p.V / 論文摘要 --- p.VII / Table of Contents --- p.IX / List of Figures --- p.XI / List of Abbreviations --- p.XIII / Chapter 1 Introduction --- p.1 / Chapter 1.1 --- Hepatocellular carcinoma (HCC) --- p.1 / Chapter 1.1.1 --- Risk factors --- p.1 / Chapter 1.1.2 --- Molecular mechanism of HCC --- p.4 / Chapter 1.1.3 --- Treatment of HCC --- p.7 / Chapter 1.2 --- Gambogic acid (GA) - a compound derived from Tradition Chinese Medicine (TCM) --- p.9 / Chapter 1.2.1 --- Traditional Chinese Medicine (TCM) --- p.9 / Chapter 1.2.2 --- Gambogic acid --- p.13 / Chapter 1.3 --- Molecular mechanism of apoptosis --- p.18 / Chapter 1.3.1 --- Overview of apoptosis --- p.18 / Chapter 1.3.2 --- Caspases cascade --- p.18 / Chapter 1.3.3 --- Bcl-2 family --- p.20 / Chapter 1.3.4 --- Mitochondria in apoptosis --- p.23 / Chapter 1.4 --- Apoptosis as a strategy for cancer therapies --- p.26 / Chapter 1.5 --- Aims of study --- p.29 / Chapter Chapter 2 --- Materials and Methods --- p.30 / Chapter 2.1 --- Cell culture and treatment --- p.30 / Chapter 2.1.1 --- Cell lines used --- p.30 / Chapter 2.1.2 --- Gambogic acid (GA) --- p.31 / Chapter 2.1.3 --- Chemicals and reagents --- p.31 / Chapter 2.1.4 --- Preparation of solutions --- p.32 / Chapter 2.1.5 --- Procedures --- p.33 / Chapter 2.2 --- Apoptotic detection --- p.35 / Chapter 2.2.1 --- Chemicals and reagents --- p.35 / Chapter 2.2.2 --- Preparation of solutions --- p.35 / Chapter 2.2.3 --- Procedures --- p.37 / Chapter 2.3 --- Effects of GA on gene expression in HepG2 --- p.41 / Chapter 2.3.1 --- Chemicals and Reagents --- p.41 / Chapter 2.3.2 --- Preparation of solutions --- p.41 / Chapter 2.3.3 --- Procedures --- p.43 / Chapter 2.4 --- Protein expression in GA-induced apoptotic cells --- p.51 / Chapter 2.4.1 --- Chemicals and Reagents --- p.51 / Chapter 2.4.2 --- Preparation of solution --- p.51 / Chapter 2.4.3 --- Procedures --- p.54 / Chapter 2.5 --- Caspase cascade study in GA-induced apoptosis --- p.60 / Chapter 2.5.1 --- Chemicals and reagents --- p.60 / Chapter 2.5.2 --- Procedures --- p.60 / Chapter 2.6 --- Downregulation of mRNA using siRNA vector --- p.62 / Chapter 2.6.1 --- siRNA expression vector --- p.62 / Chapter 2.6.2 --- Chemicals and Reagents --- p.63 / Chapter 2.6.3 --- Preparation of solution --- p.63 / Chapter 2.6.4 --- Procedures --- p.64 / Chapter Chapter 3 --- Results --- p.71 / Chapter 3.1 --- GA induces apoptosis in hepatocellular cells --- p.71 / Chapter 3.2 --- Effects of gene expression in HCC --- p.80 / Chapter 3.3 --- Caspase cascade studies in GA-induced apoptosis --- p.83 / Chapter 3.4 --- Caspase 8 activation in GA-treated cells lead to Bid cleavage --- p.89 / Chapter 3.5 --- GA induces Bax conformational changes and cytochrome c release --- p.95 / Chapter 3.6 --- Levels of protein players involved in apoptosis and cell cycle --- p.101 / Chapter Chapter 4 --- Discussion --- p.106 / References --- p.120

Thesis (Ph. D.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

Thesis (Ph. D.)--University of Hong Kong, 2010. / Includes bibliographical references (leaves 166-193). Also available in print.

Thesis (M. Med. Sc.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 71-88).

Thesis (Ph. D.)--University of Queensland, 2001. / Includes bibliographical references.

Thesis (M.D.)--University of Hong Kong, 1959. / Type written copy. Includes bibliographical references.

Thesis (M. Sc.)--University of Hong Kong, 1970. / Typeswritten.

Thesis (Ph. D.)--University of Hong Kong, 2002. / Includes bibliographical references (leaves 211-219).

Thesis (Ph. D.)--University of Queensland, 2002. / Includes bibliographical references.