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

Call number: LD2668 .T4 1950 L54 / Master of Science

As part of a health survey of New Zealand sea lions (Phocarctos hookeri) on Enderby Island, Auckland Islands (50°30’S, 166°17’E), neonatal mortality was continuously monitored at the Sandy Bay Beach rookery, from 1998/1999 to 2004/2005. The primary causes of death were categorised as trauma (35%), bacterial (24%) and hookworm (13%) infections, starvation (13%) and stillbirth (4%). During the 2001/2002 and 2002/2003 breeding seasons, bacterial epidemics caused by Klebsiella pneumoniae increased mortality by three times the mean in non-epidemic years. Uncinaria spp. from New Zealand sea lion (NZSL) pups was described for the first time using morphometric criteria. It differed from the two species already described in pinnipeds, Uncinaria lucasi and Uncinaria hamiltoni, suggesting the existence of a different morphotype in NZSLs. A study on the epidemiology of hookworm infection showed that all pups up to at least three months of age harboured adult hookworms in their intestines and transmammary transmission was identified as the route of infection of NZSL pups. Uncinariosis as a primary cause of mortality was generally associated with anaemia, haemorrhagic enteritis and frank blood in the lumen. The relationship between hookworm burden and clinical disease could not be clearly established. The 2001/2002 and 2002/2003 bacterial epidemics at Sandy Bay Beach rookery were caused by a clonal strain of Klebsiella pneumoniae as verified by pulse-field gel electrophoresis and antimicrobial testing. Suppurative arthritis was the most common post-mortem diagnosis during the two epidemic seasons. Internal lesions were consistent with septicaemia, which explained the wide range of organs from which the pathogen was grown in pure culture. A serological test investigating the exposure of NZSLs to Klebsiella spp. showed that the large majority of pups up to two months of age did not have any anti-Klebsiella antibodies, even after the epidemics, but that almost all the adults were seropositive. In addition, passive immunoglobulin (Ig) transfer from lactating females to neonates was examined by measuring IgG levels in pups and was very low compared to terrestrial mammals although similar to other pinniped neonates.

Recent advances in the field of host immunity against parasitic nematodes have revealed the importance of macrophages in trapping tissue migratory larvae. Protective immune mechanisms against the rodent hookworm Nippostrongylus brasiliensis (Nb) are mediated, at least in part, by IL-4-activated macrophages that bind and trap larvae in the lung. However, it is still not clear how host macrophages recognize the parasite. An in vitro co-culture system of bone marrow-derived macrophages and Nb infective larvae was utilized to screen for the possible ligand-receptor pair involved in macrophage attack of larvae. Competitive binding assays revealed an important role for β-glucan recognition in the process. We further identified a role for CD11b and the non-classical pattern recognition receptor ephrin-A2 (EphA2), but not the highly expressed β-glucan dectin-1 receptor, in this process of recognition. This work raises the possibility that parasitic nematodes synthesize β-glucans and it identifies CD11b and ephrin-A2 as important pattern recognition receptors involved in the host recognition of these evolutionary old pathogens. To our knowledge, this is the first time that EphA2 has been implicated in immune responses to a helminth.

Parasitische Würmer gehören mit über 3,5 Milliarden Betroffenen zu den weltweit verbreitetesten Infektionskrankheiten. Der Erfolg dieser Parasiten beruht auf ihren ausgefeilten Mechanismen mit denen sie das Immunsystem ihrer Wirte manipulieren. Interessanter Weise gehen Wurminfektionen mit einer geringeren Wahrscheinlichkeit an Allergien zu erkranken einher. Wie genau die Parasiten das Immunsystem manipulieren ist weitgehend unbekannt. Um diese Mechanismen besser studieren zu können, wurde im Rahmen dieser Arbeit versucht RNA interference (RNAi), anhand des Modellmoleküls Tropomyosin zu etablieren. Wie sich am Beispiel des Strongyliden Heligmosomoides polygyrus bakeri zeigte, ist RNAi als Manipulationsmethode für Nematoden nicht oder nur in geringem Maße geeignet. Dies lässt sich auf das Fehlen von Aufnahme- und Verbreitungsmechanismen für Doppelstrang-RNA zurückführen. Desweiteren wurden die Auswirkungen von rekombinantem Tropomyosin der Filarie Acanthocheilonema viteae (rAv-TMY) auf die Entstehung allergischer Atemwegserkrankungen im Mausmodell untersucht. Eine viermalige Behandlung mit rAv-TMY in einem Zeitraum von vier Wochen führte zu verringerten entzündlichen Reaktionen in den Atemwegen. Die Analyse immunologischer Parameter ergab, dass rAv-TMY signifikant den Einstrom von Entzündungszellen in die Atemwege reduziert, allem voran den Einstrom von Eosinophilen. Dies lässt sich durch die verringerte Ausschüttung an IL-5, Eotaxin und MCP-5 zurückführen. Zudem wurde die Bildung von antigenspezifischen IgE verringert während sich die Produktion blockierender IgG1 Antikörper erhöhte. Diese Arbeit belegt somit die anti-allergischen Eigenschaften von rAv-TMY. Damit stellt rAv-TMY ein interessantes Kandidatenmolekül zur Behandlung allergischer Reaktionen dar. Desweiteren kann der Vergleich von allergenem, nicht allergenem und modulatorischem Tropomyosin wichtige Informationen über die allgemeinen Eigenschaften von Allergenen und ihrer molekularen Struktur geben. / Parasitic worms are among the world''s most prevalent infectious diseases with more than 3.5 billion. The success of these parasites is based on their sophisticated ways to manipulate the immune system of their hosts. Interestingly, worm infections abate the risk to develop allergic disorders. How exactly parasitic worms modulate the immune system is so far largely unknown. In order to be able to investigate parasite induced modulation, this work aimed to establish RNA interference (RNAi), a method of genetic manipulation, using tropomyosin as target gene. As shown for the example of Heligmosomoides polygyrus RNAi is not or only to a small extent useful as method to genetically manipulate nematodes. This can be explained with the lack of uptake and spreading mechanisms for double stranded RNA. Furthermore, this work examined the impact of the recombinant muscle protein tropomyosin of Acanthocheilonema viteae (rAv-TMY) on the course of a rodent model of allergic airway inflammation. A four-time treatment with rAv-TMY over a period of four weeks resulted in decreased inflammatory responses in the airways. The analysis of immunological parameters showed that rAv-TMY significantly reduces the influx of inflammatory cells into the airways, especially eosinophils. The reduced eosinophil influx can be attributed to the decreased expression of IL-5, eotaxin and MCP-5 in the airways. In addition, the formation of antigen-specific IgE was impaired whereas the production of the blocking antibody IgG1 was increased. These results demonstrate the anti-allergic properties of rAv-TMY. For this reason rAv-TMY becomes an interesting model molecule for the treatment of allergic diseases. Furthermore, the comparison of allergenic, non-allergenic and modulatory tropomyosin might put some light on the nature of allergens and their molecular patterns.