Non-invasive Optical Imaging of Select Agent Bacteria in Non-human Primates

目前迫切需要开发更有效的疫苗和抗生素方案，以对抗许多导致人类致命感染的Select Agent细菌。对于候选的生物恐怖主义制剂伯克霍尔德菌来说尤其如此。假马利杆菌（Bp）和马氏伯克霍尔德菌（Bm），人类类鼻疽病和腺体的病原体。这些革兰氏阴性兼性细胞内病原体在遗传、发病机制、宿主免疫反应和临床病理等方面具有许多共同特征。马利弧菌主要感染固体，但也可通过摄入和皮肤或气溶胶暴露传染给人类。人们对这种细菌以及与之密切相关的假芽孢杆菌的担忧日益加剧，因为这种病原体似乎具有完美的特性，可以作为针对动物和人类的生物恐怖或生物战武器。在现代，它们对公共卫生的潜在破坏性影响已经升级，因为病原体在糖尿病和免疫功能低下人群中的机会性感染，这是世界上两个不断增长的人群。人类严重感染这两种病原体的死亡率都很高，它们对抗生素治疗都不耐受，而且目前还没有获得许可的疫苗用于预防或治疗。传染病的小动物模型在旨在确定可用于设计改进的诊断、预处理和针对生物恐怖制剂的治疗方法的线索和方法的研究项目中发挥着核心作用。然而，有希望的线索的进一步发展取决于更忠实地代表人类的其他动物感染模型。很明显，目前由NIAID资助的大量高质量生物防御研究将确定有希望的疾病控制新方法，而这些方法的发展将依赖于可靠、可重复和相关的感染模型。考虑到这一点，我们提出了一个u01项目，开发类鼻疽和腺体的非人类灵长类动物模型，允许直接光学成像感染。这项工作建立在我们目前的NIAID项目的基础上，该项目旨在设计兔热病和类鼻疽病的非人类灵长类动物模型。与更传统的非人类灵长类动物疾病模型相比，对感染进行成像的能力将提供一个显著的优势，因为它允许从每只动物身上收集有关感染的时间和空间格局的额外数据。此外，我们相信收集组织中细菌代谢状态的数据将是可能的。通过感染的光学成像获得的额外数据将有助于提供额外的信息，以支持临床前研究和将产品推进到人体I期临床安全性试验。此外，从每只动物获取额外数据的能力可以减少这些计划中使用的动物总数，例如通过提供来自单个动物的多个时间点数据。我们认为，开发可应用于非人类灵长类动物的敏感、非侵入性成像技术是未来临床干预选择代理细菌的重要一步。本应用程序的目的是利用全动物生物发光和荧光成像的最新进展来研究狨猴类鼻疽和腺体的发病机制和治疗。这将通过对感染动物全血的全基因组转录微阵列分析来补充，以提供对感染的免疫反应的深入看法。考虑到在封闭环境中进行的非人类灵长类动物研究的高成本和后勤限制，结合这些非致命性分析方法将提供感染动态和对治疗反应的全面图景，同时减少每项研究所需的动物数量。

There is an urgent need to develop more effective vaccines and antibiotic regimens against many of the Select Agent bacteria that cause lethal infection in humans. This is particularly true for the candidate bioterrorism agents, Burkholderia. pseudomallei (Bp) and Burkholderia mallei (Bm), the causative agents of human melioidosis and glanders. These Gram negative, facultative intracellular pathogens share many common features of their genetics, pathogenesis, host immune response and clinical pathology. B. mallei primarily infect solipeds but the disease is transmissible to humans by ingestion and cutaneous or aerosol exposures. Concern over this bacterium and the very closely related species B. pseudomallei has heightened because of the pathogens' seemingly perfect characteristics for malevolent uses as bioterror or biowarfare weapons against both animals and humans. In modern times their potential destructive impact on public health has escalated due to the pathogens' opportunistic infection of diabetic and immunocompromised people, two growing populations worldwide. For both pathogens, severe infection in humans carries a high mortality rate, all are recalcitrant to antibiotic therapy and no licensed vaccine exists for either prophylactic or therapeutic use. Small animal models of infectious disease play a central role in research programs aimed at identifying leads and approaches that could be exploited to devise improved diagnostics, pre-treatments and therapies against bioterrorism agents. However, the further development of promising leads is dependent on additional animal models of infection which more faithfully represent humans. It is clear that the large body of high quality biodefense research currently funded by NIAID will identify promising new approaches to disease control, and that the development of these approaches will be dependent on reliable, reproducible and relevant models of infection. With this in mind we propose a UO1 project to develop non-human primate models of melioidosis and glanders which allow the direct optical imaging of the infection. This work builds on our current NIAID projects to devise non-human primate models of tularaemia and melioidosis. The ability to image the infection will provide a significant advantage over more conventional non-human primate models of disease, because it allows additional data on the temporal and spatial pattern of the infection to be gathered from each animal. In addition, we believe it will be possible to gather data on the metabolic status of the bacteria in tissues. The additional data acquired by optical imaging of the infection will have benefits in providing additional information to support preclinical studies and advancement of products into phase I clinical safety trials in humans. Also, the ability to acquire additional data from each animal could reduce the overall numbers of animal used in these programmes, for example by providing multiple time point data from a single animal. We believe that the development of sensitive, non-invasive imaging techniques which can be applied to non human primates is an essential step in the generation of future clinical interventions to Select Agent bacteria. The purpose of this application is to apply recent advances in whole animal bioluminescent and fluorescent imaging to study the pathogenesis and treatment of melioidosis and glanders in marmosets. This will be complemented by genome-wide transcriptional microarray analysis of whole blood from infected animals to provide an in-depth view of the immunological response to infection. Given the high cost and logistical constraints of non-human primate studies conducted in containment, combining these non- lethal analytical approaches will provide a comprehensive picture of the dynamics of infection and response to treatment, while reducing the number of animals needed for each study. PHS 398/2650 (Rev. 11/07) Page 12 Continuation Format Page