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，以及细胞间的信号传递。了解这些将使我们更好地理解如何从药理学、免疫学和遗传学上操纵免疫系统。我们有一个复杂的投资组合，涉及患者、动物和实验室标本。我们跟踪了大量NADPH氧化酶缺乏的患者，慢性肉芽肿病(CGD)，我们一直参与描述他们的感染和他们发展的并发症。我们还使用了我实验室培育的一只小鼠，它缺乏NADPH氧化酶，因此与人类的CGD非常相似。在这些小鼠身上的大量研究表明，这种酶系统不仅在保护免受感染方面发挥着关键作用，而且在炎症反应的大小和特征方面也发挥着关键作用。这种混合的方法来理解CGD中的NADPH氧化酶，对于先天性免疫系统在炎症反应的早期和晚期所起的作用非常有意义。 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，即致密性颗粒状杆菌。这种细菌已经被注射到CGD的动物模型中，并被证明是CGD的重要病原体。我们目前已在CGD患者中发现了5例球虫感染，表明这是CGD中的一种重要感染。由血清学确定，这种感染在正常人中的发生率很高。这种联合的长凳和床边治疗CGD的方法将使我们能够识别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，并更好地了解炎症。 与合作者在小鼠模型中合作，我们已经确定CGD病原体与CGD细胞的相互作用不同于非病原体，这为我们提供了一套探索了解CGD毒力的途径。今年，我们在CGD中发现了几种新的真菌感染，包括新沙里氏杆菌，并在CGD和HIE中发现了新的带有绿色曲霉的综合征表现。 高IgE反复感染综合征(HIEs或约伯氏综合征)的遗传学和细胞学基础的鉴定，这是一种常染色体显性疾病，特征是IgE极度升高，反复的中国-肺部感染，骨量减少，脊柱后凸，肺囊肿和牙齿异常，因为STAT3激活了广泛的研究领域。根据临床证据，我们知道约伯的基因对先天免疫、早期和晚期宿主免疫反应、骨骼生长发育和牙齿蜕膜肯定是至关重要的。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-表型相关性。与NIAID和NIAMS的合作者一起，我们发现了STAT3下游的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。 这些结合起来的方法继续是富有成效的，并将导致更好地理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 let us better appreciate 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 the NADPH oxidase deficiency, 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 this enzyme system 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 5 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 inflammation better. Working with collaborators in the mouse model we have determined that CGD pathogens interact with CGD cells diferntly than do non-pathogens, giving us a set of pathways to explore to understand CGD virulence. We have identified several novel fungal infections in CGD this year, including Neosartorya udagawae, and identified novel syndromic presentations with Aspergillus viridinutans in CGD and HIES. 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. 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.