Identification of novel innate immune mechanisms regulating inflammation in the gastrointestinal tract

The Gastro-Intestinal (GI) tract is a unique region in the body. Our innate immune system retains a fine homeostatic balance between avoiding inappropriate inflammatory responses against the myriad commensal microbes residing in the gut while also remaining active enough to prevent invasive pathogenic attack. The intestinal epithelium represents the frontline of this interface. It has long been known to act as a physical barrier preventing the lumenal bacteria of the gastro-intestinal tract from activating an inflammatory immune response in the immune cells of the underlying mucosa. However, in recent years, an appreciation has grown surrounding the role played by the intestinal epithelium in regulating innate immune responses, both in the prevention of infection and in maintaining a homeostatic environment through modulation of innate immune signalling systems. The aim of this thesis was to identify novel innate immune mechanisms regulating inflammation in the GI tract. To achieve this aim, we chose several aspects of regulatory mechanisms utilised in this region by the innate immune system. We identified several commensal strains of bacteria expressing proteins containing signalling domains used by Pattern Recognition Receptors (PRRs) of the innate immune system. Three such bacterial proteins were studied for their potentially subversive roles in host innate immune signalling as a means of regulating homeostasis in the GI tract. We also examined differential responses to PRR activation depending on their sub-cellular localisation. This was investigated based on reports that apical Toll-Like Receptor (TLR) 9 activation resulted in abrogation of inflammatory responses mediated by other TLRs in Intestinal Epithelial Cells (IECs) such as basolateral TLR4 activation. Using the well-studied invasive intra-cellular pathogen Listeria monocytogenes as a model for infection, we also used a PRR siRNA library screening technique to identify novel PRRs used by IECs in both inhibition and activation of inflammatory responses. Many of the PRRs identified in this screen were previously believed not to be expressed in IECs. Furthermore, the same study has led to the identification of the previously uncharacterised TLR10 as a functional inflammatory receptor of IECs. Further analysis revealed a similar role in macrophages where it was shown to respond to intracellular and motile pathogens such as Gram-positive L.monocytogenes and Gram negative Salmonella typhimurium. TLR10 expression in IECs was predominantly intracellular. This is likely in order to avoid inappropriate inflammatory activation through the recognition of commensal microbial antigens on the apical cell surface of IECs. Moreover, these results have revealed a more complex network of innate immune signalling mechanisms involved in both activating and inhibiting inflammatory responses in IECs than was previously believed. This contribution to our understanding of innate immune regulation in this region has several direct and indirect benefits. The identification of several novel PRRs involved in activating and inhibiting inflammation in the GI tract may be used as novel therapeutic targets in the treatment of disease; both for inducing tolerance and reducing inflammation, or indeed, as targets for adjuvant activation in the development of oral vaccines against pathogenic attack.

Nietzsche and Montaigne: cheerful naturalism

The influence of the Essays of Michel de Montaigne on the thought of Friedrich Nietzsche has, hitherto, received scant scholarly attention. The aim of this thesis is to address this lacuna in the literature by making evident the importance of the Essays to the development of Nietzsche’s philosophy. I argue that, in order to fully appreciate Nietzsche’s thought, it must be recognized that, from the beginning to the end of his philosophical life, Montaigne was for him a thinker of the deepest personal and philosophical significance. Against the received scholarly opinion, which would see Montaigne as influential only for Nietzsche’s middle works, I contend that the Essays continue to be a key inspiration for Nietzsche even into his late and final works. Montaigne, with his cheerful affirmation of life, his experimental mode of philosophizing, and his resolutely naturalized perspective, was an exemplar for Nietzsche as a philosopher, psychologist, sceptic and naturalist. The Essays not only stimulated Nietzsche’s thinking on questions to do with morality, epistemology and the nature of the soul but also informed his conception of the ideal philosophical life. Moreover, to explore the Essays from a Nietzschean viewpoint, allows the drawing out of the more radical aspects of Montaigne’s thought, while to probe Montaigne’s impact on Nietzsche, provides insight into the trajectory of Nietzsche’s philosophy as he broke free from romantic pessimism and embraced the naturalism that would guide his works from Human, All Too Human onward.