Transgenic Animal Models Of Human Immune Defects

人类免疫缺陷的转基因动物模型

• 批准号: 8156872
• 负责人: Steven Holland
• 金额: $ 201.65万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8156872
• 关键词: Animal Model; Defect; Human; Immune; Transgenic Animals

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.

我们寻求了解酶系统（特别是 NADPH 氧化酶）在正常宿主功能和免疫防御中的作用。 NADPH 氧化酶参与炎症的产生和控制、感染保护以及细胞间信号传导。了解这些将告诉我们如何从药理学、免疫学和遗传学上操纵免疫系统。我们拥有复杂的产品组合，涉及患者、动物和实验室标本。我们跟踪了大量患有慢性肉芽肿病 (CGD) 的患者，并参与了他们的感染和并发症的特征分析。我们还使用了实验室培育的小鼠，该小鼠缺乏 NADPH 氧化酶，因此与人类 CGD 非常相似。对这些小鼠的大量研究表明，NADPH 氧化酶不仅在防止感染方面发挥着关键作用，而且在炎症反应的程度和特征方面也发挥着关键作用。这种了解 CGD 中 NADPH 氧化酶的混合方法为了解先天免疫系统在炎症反应的早期和晚期方面的作用提供了非常丰富的信息。 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，即贝塞斯登颗粒杆菌 (Granulibacter bethesdensis)。该微生物已被注射到 CGD 动物模型中，并被证明是 CGD 的重要病原体。我们现在在CGD患者中总共发现了6例G. bethesdensis，表明这是CGD的重要感染。根据血清学测定，正常人中这种感染的频率很高。这种结合临床和床边的 CGD 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，并了解和治疗 CGD 炎症。 通过与小鼠模型中的合作者合作，我们确定了 CGD 病原体与 CGD 细胞的相互作用与非病原体不同，这为我们提供了一组探索了解 CGD 毒力的途径。我们已经发现了 CGD 中的几种新感染，包括酸单胞菌和甲基杆菌，它们与颗粒杆菌相一致，表明甲基营养生物在 CGD 中发挥着作用。 高 IgE 复发性感染综合征（HIES 或乔布氏综合征）是一种常染色体显性遗传疾病，其特征是 IgE 极度升高、复发性肺部感染、骨质减少、脊柱后侧凸、肺囊肿和牙齿异常，鉴定其遗传和细胞基础，因为 STAT3 已激活了广泛的研究领域。从临床角度来看，我们知道乔布氏病涉及的基因对于先天免疫、早期和晚期宿主免疫反应、骨骼生长和发育以及牙齿脱落至关重要。我们在体外重建了 STAT3 突变，以研究它们的排列和可能的基因型-表型相关性。我们与 NIAID 和 NIAMS 合作者一起发现了 STAT3 下游其他细胞因子的异常，最明显的是 IL-17，它在乔布斯综合征患者的细胞中含量极低。我们与 NIAMS 的研究人员合作创建了 STAT3 缺陷的小鼠模型。我们与 NHLBI 的研究人员合作，在体外研究了 STAT3 缺陷患者的血管内皮细胞。 这些组合方法将继续发挥作用，并将有助于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。