Transgenic Animal Models Of Human Immune Defects

我们试图了解酶系统在正常宿主功能和免疫防御中的作用，特别是NADPH氧化酶。NADPH氧化酶参与炎症的产生和控制，保护免受感染和细胞-细胞信号传导。了解这些将告诉我们如何操纵免疫系统，免疫学和遗传学。我们有一个复杂的产品组合，涉及患者，动物和实验室标本。我们随访了大量慢性肉芽肿病（CGD）患者，并参与了描述其感染和并发症的研究。我们还使用了在我的实验室中创建的小鼠，该小鼠缺乏NADPH氧化酶，因此与人类CGD非常相似。在这些小鼠中进行的大量研究表明，NADPH氧化酶不仅在保护免受感染方面，而且在炎症反应的程度和特征方面发挥关键作用。这种理解CGD中NADPH氧化酶的混合方法对于先天免疫系统在炎症反应的早期和晚期方面的作用非常有用。 通过对CGD患者的研究，我们发现了一种引起CGD感染的新细菌，贝氏颗粒杆菌。该生物体已被注射到CGD的动物模型中，并显示为CGD中的重要病原体。我们现在已经确定了6例G。Bethesdensis在CGD患者中，表明这是CGD中的重要感染。通过血清学测定，这种感染在正常人中的频率很高。这种联合的实验室和床旁CGD方法将使我们能够确定宿主和细菌生理学的重要方面，这将有助于我们更好地治疗CGD患者，并了解和治疗CGD炎症。 在小鼠模型中与合作者合作，我们已经确定CGD病原体与CGD细胞的相互作用与非病原体不同，这为我们提供了一系列探索了解CGD毒力的途径。我们已经确定了几个新的感染CGD包括嗜酸杆菌和甲基杆菌，在copncert与颗粒杆菌，表明甲基营养生物在CGD中的作用。 高IgE复发性感染综合征（HIES或Job综合征）的遗传和细胞基础的鉴定，这是一种常染色体显性遗传疾病，其特征是IgE极高、复发性鼻窦-肺感染、骨质减少、脊柱后凸、肺囊肿和牙齿异常，因为STAT 3已经激活了广泛的研究领域。从临床的基础上，我们知道，基因参与的工作必须是至关重要的先天免疫，早期和晚期的宿主免疫反应，骨骼的生长和发育，牙齿脱落。我们已经在体外重建了STAT3突变，以研究它们的排列和可能的基因型-表型相关性。通过NIAID和NIAMS的合作，我们已经确定了STAT3下游其他细胞因子的异常，最明显的是IL-17，它在约伯综合征患者的细胞中非常低。与NIAMS的研究人员合作，我们创建了STAT3缺陷的小鼠模型。与NHLBI的研究人员合作，我们在体外研究了STAT3缺陷患者的血管内皮细胞。 这些结合的方法继续是富有成效的，并将导致更好地了解先天免疫和炎症的途径。我们相信这些研究将帮助我们在分子遗传和功能水平上了解几种不同的感染，包括真菌感染。

We seek to understand the role of enzyme systems in normal host function and immune defense, specifically the NADPH oxidase. The NADPH oxidase is involved in the generation and control of inflammation, protection from infection, and cell-cell signaling. Understanding these will inform us about how to manipulate the immune system pharmacologically, immunologically, and genetically. We have a complex portfolio involving patients, animals, and laboratory specimens. We follow a large number of patients with chronic granulomatous disease (CGD), and we have been involved in characterizing their infections and the complications they develop. We have also used a mouse created in my laboratory that is deficient in the NADPH oxidase and therefore closely mimics human CGD. Numerous studies in these mice have shown a critical role for the NADPH oxidase in not only protection from infection but also in the magnitude and character of the inflammatory response. This mixed approach to understanding the NADPH oxidase in CGD has been very informative about the role of the innate immune system in both early and late aspects of the inflammatory response. Through the study of patients with CGD we have identified a new bacteria that causes infection in CGD, Granulibacter bethesdensis. This organism has been injected into animal models of CGD and shown to be a significant pathogen in CGD. We have now identified a total of 6 cases of G. bethesdensis in CGD patients, indicating that this is an important infection in CGD. The frequency of this infection in normals as determined by serology is high. This conjoined bench and bedside approach to CGD will allow us to identify important aspects of both host and bacterial physiology that will help us treat patients with CGD better, and to understand and treat CGD inflammation. Working with collaborators in the mouse model we have determined that CGD pathogens interact with CGD cells differently than do non-pathogens, giving us a set of pathways to explore to understand CGD virulence. We have identified several novel infections in CGD including Acidomonas and Methylobacter, which, in copncert with Granulibacter, suggest a role for methylotrophic organisms in CGD. Identification of the genetic and cellular basis of hyper-IgE recurrent infection syndrome (HIES or Job's syndrome), an autosomal dominant disease characterized by extremely elevated IgE, recurrent sino-pulmonary infections, osteopenia, kyphoscoliosis, pulmonary cysts, and dental abnormalities, as STAT3 has activated broad areas of investigation. From clinical grounds we knew that the gene involved in Job's must be critically important to innate immunity, the early and late host immune responses, skeletal growth and development, and tooth deciduation. We have recreated STAT3 mutations in vitro to study their permutations and possible genotype-phenotype correlations. With NIAID and NIAMS collaborators we have identified abnormalities in other cytokines downstream of STAT3, most notably IL-17, which is profoundly low in cells from Job's syndrome patients. Collaborating with investigators in NIAMS we have created a mouse model of STAT3 deficiency. Collaborating with investigators in NHLBI we have studied vascular endothelial cells from patients with STAT3 deficeincy in vitro. These combined approaches continue to be productive and will lead to better understanding of the pathways of innate immunity and inflammation. We believe that these studies will help us understand several different infections, including fungal infections, at a molecular genetic and functional level.