Development of multi-parametric measurements of the circulating immune response to Feline Tuberculosis to improve the understanding and diagnosis of t

结核病(TB)是人类和动物的一个主要全球性问题。在英国，结核病在牛、獾和包括宠物猫在内的其他家养和野生动物物种中是一个日益严重的问题。我们已经表明，英国的猫有很大的结核病风险，因为大约1%的常规猫组织活检具有与分枝杆菌病一致的组织病理学变化(Gunn-Moore等人。2013年)；其中至少15%是由于牛分枝杆菌，19%是微小分枝杆菌(田鼠/啮齿动物杆菌)(Gunn-Moore等人)。2011年)。猫中的一些牛分枝杆菌感染进展极快，感染组织和渗出物中存在大量微生物；鉴于最近认识到猫对人类人畜共患病的风险，这构成了一个主要关切(Roberts等人)。2014年)。我们假设，家猫接触牛分枝杆菌或微小分枝杆菌的结果以及人畜共患传播的可能性，是由病原体的性质和宿主的免疫反应决定的。快速、准确的诊断是确定人畜共患病风险和管理方案的关键。目前，猫的结核病是通过培养、干扰素-伽马(干扰素-伽马)释放试验(IGRA)或聚合酶链式反应(PCR)试验来确认的。然而，这三种方法都有很大的局限性：高达50%的感染样本无法培养(尽管存在ZN+分枝杆菌)；IGRA不能总是区分牛分枝杆菌和微小分枝杆菌；而且在可用的情况下，PCR检测非常昂贵。这项研究旨在改进猫结核病的诊断测试，这将对确定猫的人畜共患病以及牛分枝杆菌和微小分枝杆菌感染的物种间和物种内风险产生重大影响。它还将为管理选择提供信息，预测生存(清除或潜伏的可能性)，从而对受感染猫、它们的主人以及接触过的家养和野生动物物种的长期健康产生影响。一个重要的成果将是更好地了解家猫对牛分枝杆菌或微小分枝杆菌感染的免疫反应。该项目建立在对感染结核病的猫的免疫反应正在进行的研究的基础上。我们有一个重要的血清、血浆和福尔马林固定的石蜡包埋组织(&gt；300)样本银行，我们将继续扩大这一收集范围。利用这些样本，我们最近确定了感染牛分枝杆菌或微小分枝杆菌的猫的细胞因子谱，并显示出感染和健康猫之间，以及牛分枝杆菌和微小分枝杆菌感染之间的显著差异。在平行研究中，我们还显示了感染猫的巨噬细胞反应和肉芽肿炎症的显着差异，这是感染性暴露(准备中的多发性硬化症)结局的重要决定因素。与我们的工业合作伙伴Biobest一起，我们正在继续评估IGRA测试的敏感性和特异性。这一新的PHD项目将以四个主要目标扩展这些研究：1)继续从结核猫和健康对照中收集样本，以扩大我们的生物库，并为该项目的分析提供额外的样本。2)使用多重细胞因子分析来测量血清/血浆细胞因子，以确定测量额外的细胞因子(与干扰素γ一起)是否可以提高诊断的敏感性和特异性。这也将告知对感染牛分枝杆菌和微小分枝杆菌的免疫反应。3)与行业合作伙伴合作开发多重抗体检测系统，以阐明宿主体液免疫反应对感染的作用。4)将(2)和(3)的结果与临床病史、结果和治疗反应相关联，以确定可以预测致病程度、传播风险和可能的治疗反应(在适当情况下)的特征图谱。该项目将使学生获得广泛的技能专业知识，并将与Biobest进行广泛的合作，Biobest将成为这项工作的行业合作伙伴。

Tuberculosis (Tb) is a major global concern in humans and animals. In the UK, Tb is an increasing problem in cattle, badger and other domestic and wildlife species, including pet cats. We have shown that cats in the UK are significantly at risk of Tb, as ~1% of all routine feline tissue biopsies have histopathological changes consistent with mycobacteriosis (Gunn-Moore et al. 2013); at least 15% of these are due to M. bovis, and 19% M. microti (the vole/rodent bacillus) (Gunn-Moore et al. 2011). Some M. bovis infections in cats progress extremely rapidly, with large numbers of organisms present within the affected tissues and exudates; this poses a major concern given the recently recognised cat to human zoonotic risk (Roberts et al. 2014). We hypothesise that the outcome of exposure of domestic cats to M. bovis or M. microti, and the likelihood of zoonotic transmission, is determined by the nature of the pathogen and the host's immune response.Rapid, accurate, diagnosis is the key to determining zoonotic risk and management options. Currently, Tb in cats is confirmed by culture, IFN-gamma (IFN gamma) release assay (IGRA) or PCR tests. However, there are major limitations with all three methodologies: infected samples fail to culture in up to 50% of cases (despite having ZN+ mycobacteria present); the IGRA cannot always differentiate M. bovis from M. microti; and PCR tests, where available, are very costly. This study aims to improve diagnostic tests for feline Tb that will impact significantly on determining the zoonotic and inter- and intra-species risk of M. bovis and M. microti infections in cats. It will also inform management options, predict survival (likelihood of clearance or latency), and so impact on the long term health of infected cats, their owners, and in-contact domestic and wildlife species. An important output will be a greater understanding of the immune response of domestic cats to infection with M. bovis or M. microti. The project builds upon ongoing studies of the immune response of cats infected with Tb. We have in place a significant bank of samples of serum, plasma and formalin fixed paraffin-embedded tissues (>300) from Tb cats and we will continue to expand this collection. Using these samples we have recently defined the cytokine profile of cats infected with M. bovis or M. microti and shown significant differences between infected and healthy cats, and between M. bovis and M. microti infection. In parallel studies we have also shown significant differences in macrophage responses and granulomatous inflammation in infected cats which are important determinants in the outcome of infectious exposure (MS in preparation). With our industrial partner Biobest we are continuing to evaluate the sensitivity and specificity of the IGRA test. This new PhD project will expand these studies with four major objectives:1) Continue to collect samples from tuberculous cats and healthy controls to expand our biobank and provide additional samples for analysis within this project.2) Measure serum/plasma cytokines using a multiplex cytokine assay to determine whether measurement of additional cytokines (alongside IFN gamma) can improve diagnostic sensitivity and specificity. This will also inform on the immune response to infection with M. bovis and M. microti. 3) Develop, in collaboration with industrial partners, a multiplex antibody detection system that will elucidate the role of the host's humoral immune response to infection.4) Correlate the outcome of (2) and (3) with clinical histories, outcomes and responses to treatment to identify signature profiles that can predict the degree of pathogenicity, the risk of transmission, and the likely response to treatment (where appropriate).The project will enable the student to gain expertise in a wide range of skills and will involve extensive collaboration with Biobest who will be the industrial partner for this work.