Thesis (Ph. D.)--University of Texas at Austin, 2005. / Supervisor: Jeanne H. Freeland-Graves. Vita. Includes bibliographical references.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Thesis (M. Med. Sc.)--University of Hong Kong, 2004. / Also available in print.

by Liu Heng. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 115-138). / Acknowledgments --- p.I / List of Abbreviations --- p.II / Abstract --- p.V / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Cytokines in the Central Nervous System --- p.1 / Chapter 1.1.1 --- Basic Properties of Cytokines --- p.1 / Chapter 1.1.2 --- The General Characteristics of Glial Cells --- p.4 / Chapter 1.1.2.1 --- Astrocytes --- p.4 / Chapter 1.1.2.2 --- Oligodendrocytes --- p.6 / Chapter 1.1.2.3 --- Microglial --- p.7 / Chapter 1.1.3 --- The Effects of Cytokines on Neural Cells --- p.7 / Chapter 1.1.3.1 --- TNF-α and Neural Cells --- p.8 / Chapter 1.1.3.2 --- LIF and Neural Cells --- p.10 / Chapter 1.1.3.3 --- IL-1 and Neural Cells --- p.12 / Chapter 1.1.3.4 --- IL-6 and Neural Cells --- p.14 / Chapter 1.1.4 --- Immune Response in the Central Nervous System --- p.16 / Chapter 1.2 --- The C6 Glioma as a Model for the Study of Glial Cell Growth and Differentiation --- p.21 / Chapter 1.2.1 --- The Rat C6 Glioma Cells --- p.21 / Chapter 1.2.2 --- The Differentiation and Proliferation of C6 Glioma Cells --- p.23 / Chapter 1.3 --- Signal Transduction Pathways in Cytokine-stimulated Glial Cells --- p.28 / Chapter 1.3.1 --- Intracellular Signalling Pathways of Cytokines --- p.28 / Chapter 1.3.1.1 --- Protein Kinase C Pathway --- p.29 / Chapter 1.3.1.2 --- Tyrosine Kinase Pathway --- p.30 / Chapter 1.3.1.3 --- Cyclic Nucleotide Pathway --- p.32 / Chapter 1.3.1.4 --- Nitric Oxide Pathway --- p.33 / Chapter 1.3.2 --- Intracellular Signalling Pathways in Cytokine-stimulated C6 Glioma Cells --- p.34 / Chapter 1.4 --- The Aims of This Thesis Project --- p.37 / Chapter Chapter 2: --- Materials and Methods --- p.41 / Chapter 2.1 --- Rat C6 Glioma Cell Culture --- p.41 / Chapter 2.1.1 --- Preparation of Culture Media --- p.41 / Chapter 2.1.1.1 --- Complete Dulbecco's Modified Eagle Medium --- p.41 / Chapter 2.1.1.2 --- Complete Roswell Park Memorial Institute1640 Medium --- p.42 / Chapter 2.1.2 --- Maintenance of the C6 Cell Line --- p.42 / Chapter 2.1.3 --- Cell Preparation for Assays --- p.43 / Chapter 2.2 --- Determination of Cell Proliferation --- p.44 / Chapter 2.2.1 --- Determination of Cell Proliferation by [3H]-Thymidine Incorporation --- p.44 / Chapter 2.2.2 --- Measurement of Cell Viability Using Neutral Red Assay --- p.45 / Chapter 2.2.3 --- Data Analysis --- p.45 / Chapter 2.3 --- Effects of Cytokines and Lipopolysaccharide on C6 Cell Proliferation --- p.46 / Chapter 2.4 --- Effects of Protein Kinase C Activators and Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.47 / Chapter 2.5 --- Effects of cAMP or cGMP on Cytokine-induced C6 Cell Proliferation --- p.48 / Chapter 2.6 --- Effects of Tyrosine Kinase Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.48 / Chapter 2.7 --- Effects of Calcium Ion on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8 --- Effects of Nitric Oxide on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8.1 --- Effects of Sodium Nitroprusside and Nitric Oxide Synthase Inhibitors on Cytokine-induced C6 Cell Proliferation --- p.49 / Chapter 2.8.2 --- Nitric Oxide Production Assay --- p.50 / Chapter 2.9 --- Effects of β-Adrenergic Receptor Agonist and Antagonist on Cytokine-induced C6 Cell Proliferation --- p.51 / Chapter 2.10 --- Morphological Studies on Cytokine-Treated C6 Glioma Cells --- p.51 / Chapter 2.10.1 --- Wright-Giesma Staining --- p.52 / Chapter 2.10.2 --- Glial Fibrillary Acidic Protein Staining --- p.52 / Chapter 2.10.3 --- Hematoxylin Staining --- p.53 / Chapter Chapter 3: --- Results --- p.55 / Chapter 3.1 --- Effects of Cytokines on C6 Cell Proliferation --- p.55 / Chapter 3.1.1 --- Effects of Cytokines on C6 Cell Proliferation --- p.56 / Chapter 3.1.2 --- The Time Course of Cytokine-induced C6 Cell Proliferation --- p.59 / Chapter 3.1.3 --- Effects of Lipopolysaccharide on C6 Cell Proliferation --- p.61 / Chapter 3.1.4 --- Effects of Cytokines on the Growth of C6 Cells --- p.64 / Chapter 3.2 --- Morphology and GFAP Expression in Cytokine-treated C6 Glioma Cells --- p.64 / Chapter 3.2.1 --- Effects of Cytokines on the Morphology of C6 Cells --- p.64 / Chapter 3.2.2 --- Effects of Cytokines on GFAP Expression in C6 Glioma Cells --- p.66 / Chapter 3.3 --- The Signalling Pathway of Cytokine-induced C6 Cell Proliferation --- p.69 / Chapter 3.3.1 --- The Involvement of Protein Kinase C in Cytokine-induced C6Cell Proliferation --- p.71 / Chapter 3.3.2 --- The Involvement of Tyrosine Kinase in the Cytokine- induced C6 Cell Proliferation --- p.81 / Chapter 3.3.3 --- The Involvement of Calcium Ions in Cytokine-induced C6 Cell Proliferation --- p.87 / Chapter 3.3.4 --- The Involvement of Cyclic Nucleotides in Cytokine- induced C6 Cell Proliferation --- p.92 / Chapter 3.3.5 --- The Involvement of Nitric Oxide in Cytokine-induced C6 Cell proliferation --- p.94 / Chapter 3.3.6 --- The Involvement of P-Adrenergic Receptor in Cytokine- induced C6 Cell Proliferation --- p.101 / Chapter Chapter 4: --- Discussion and Conclusions --- p.104 / References --- p.115

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Thesis (M.D.)--University of Adelaide, Faculty of Medicine, 1994. / Includes bibliographical references (leaves 152-193).

Having passed all pre-clinical safety testing, the superagonistic anti -CD28 therapeutic monoclonal antibody (mAb ) TGN 1412, intended for the treatment of rheumatoid arthritis and B-cell chronic lymphocytic leukaemia, was approved by German and UK regulatory authorities for first-in-man Phase One clinical trial. Shortly after infusion, all six healthy trial volunteers suffered unexpected and profound systemic pro-inflammatory cytokine release, later termed a 'cytokine storm,' causing multi-organ failure. This unexpected and near fatal cytokine release syndrome (CRS) publically highlighted the failure of current pre-clinical safety testing procedures, emphasising an urgent need for novel cytokine release assays (CRAs) capable of predicting adverse properties of therapeutic mAbs. A wet coat mAb immobilisation approach, developed here, has proven predictive of clinical outcome and would have anticipated TGN1412 immunotoxicity in man. This approach IS now being widely applied by the pharmaceutical industry and contract research organisations (CROs). Comparative studies, testing TGN 1412 against a panel , of therapeutic mAbs, identified a unique mechanism of TGNl412-driven cytokine release. Substantial concentrations of the cytokine lL-2 were subsequently found to be a hallmark for the cytokine storm observed in-vivo, signifYing IL-2 as a TGN1412-like response biomarker. Multiple pro-inflammatory cytokine release was also shown to be principally effector memory T cell (T EM) derived in man. Human and macaque comparative immunophenotyping crucially identified macaque T EM cells as lacking CD28 expression, explaining pre-clinical animal model testing failures. A more physiologically relevant aqueous phase co-culture assay using monocyte-derived dendritic cells is also shown capable of eliciting a TGN1412-like cytokine release profile equivalent to that detected in-vivo and in-vitro using wet coat immobilisation; implying presentation in-vivo likely involved dendritic cells. This thesis describes the most likely mechanisms of action responsible for TGN1412 immunotoxicology in man and provides a plausible explanation for the pre-clinical safety testing failures, findings vital to the fields of immunomodulatory therapeutic mAb development and immunotoxicology.

No description available.

published_or_final_version / Obstetrics and Gynaecology / Master / Master of Medical Sciences

published_or_final_version / Medicine / Master / Master of Research in Medicine