Transgenic Animal Models Of Human Immune Defects

我们试图通过检查免疫缺陷的后果来了解吞噬细胞在免疫功能中的作用。我们的主要关注点是慢性肉芽肿性疾病(CGD)，它是由NADPH氧化酶缺陷引起的。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。我们有一个全面的产品组合，涉及患者、动物和实验室标本。 在过去的一年里，我们在描述残余NADPH活性在CGD患者生存中的作用方面进行了合作(Kuhns等人，NEJM 2010)。本文阐述了超氧化物在感染易感性和生存中的作用。令人惊讶的是，研究表明，超氧化物的产生与炎症性肠病无关，但与肝病有关。 我们还发表了一篇关于阻断肿瘤坏死因子在CGD相关炎症性肠病中的并发症，导致严重感染和死亡的开创性论文(Uzel等人，CID 2010)。这是关于残留的肿瘤坏死因子在CGD预防感染中的作用的第一份报告，并对CGD中使用这些药物提出了重要的警告。有趣的是，所有CGD患者都有与炎症性肠病相关的抗体水平升高，无论是有肠道主诉还是没有肠道主诉的患者(Yu等人，ClinImm 2011)。因此，这些血清学检测在CGD中没有价值。 我们继续探索致密性颗粒状杆菌的血清流行病学，这是一种新的革兰氏阴性菌，宿主范围仅限于CGD。我们已经证实CGD患者的血清阳性率几乎为75%，但在正常人中的血清阳性率高达25%。 高IgE反复感染综合征(HIEs或约伯氏综合征)的遗传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异常升高，反复发生的中国-肺部感染，骨量减少，脊柱后凸，肺囊肿和牙齿异常，因为STAT3提供了广泛的研究领域。通过NIAID、NIAMS和校外合作者，我们已经发现STAT3下游的其他细胞因子异常，最明显的是IL-17，它在约伯综合征患者的细胞中非常低。我们最近发现，唾液中的IL-17水平在杀死口腔念珠菌和STAT3缺乏症严重受损的人方面起着重要作用。与NIAMS的研究人员合作，我们创建了STAT3缺乏症的小鼠模型，这种缺乏症损害了伤口愈合和葡萄球菌控制。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，从表现出趋化因子产生受损的STAT3缺陷患者的血管内皮细胞和肌肉细胞中分离出来。 最近我们检查了一例轻度中性粒细胞减少、血小板减少、反复感染和严重影响口腔伤口愈合的患者。通过仔细的生物化学和分子方法，我们已经确定这名儿童在支架蛋白WDR1中有一种新的缺陷。足够有趣的是，我们在25年前看到的一个患有类似病理的儿童身上发现了同样的缺陷，并确定了同一基因的突变。 这些结合起来的方法仍然是有效的，并帮助我们了解先天免疫和炎症。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 phagocytes in immune function through examination of the consequences of immune defects. Our major focus is on chronic granulomatous disease (CGD), which is caused by defects in the enzyme NADPH oxidase. The NADPH oxidase is involved in the generation and control of inflammation, protection from infection, and cell-cell signaling. We have a comprehensive portfolio involving patients, animals, and laboratory specimens. Over the last year we have collaborated in the characterization of the role of residual NADPH activity in survival of CGD patients (Kuhns et al, NEJM 2010). This paper clarified the role of superoxide in infection susceptibility and survival. Surprisingly, it showed that superoxide production is not tied to inflammatory bowel disease but is related to liver disease. We also published a seminal paper on the complications of TNF blockade in CGD-related inflammatory bowel disease, leading to severe infections and death (Uzel et al, CID 2010). This is the first report of the role of residual TNF in protection from infection in CGD and a significant caution for the use of these drugs in CGD. Interestingly, all patients with CGD have elevated levels of antibodies associated with inflammatory bowel disease, both those with and without bowel complaints (Yu et al, ClinImm 2011). Therefore, these serologic tests are of no value in CGD. We have continued our exploration of the seroepidemiology of Granulibacter bethesdensis, a novel Gram negative organism with a host range limited to CGD. We have confirmed a seropositivity rate of almost 75% in CGD patients, but an impressive 25% in normal individuals. 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 informed broad areas of investigation. With NIAID, NIAMS and extramural 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. We have recently shown that salivary levels of IL-17 are important in killing of oral candida and serioulsy impaired in STAT3 deficient humans. Collaborating with investigators in NIAMS we have created a mouse model of STAT3 deficiency which has impaired wound healing and staphylococcal control. Collaborating with investigators in NHLBI we have studied vascular endothelial cells from patients with STAT3 deficeincy in vitro, deriving endothelial and muscle cells from STAT3 deficient patients that have shown impaired chemokine production. Recently we examined a patient with mild neutropenia, thrombocytopenia, recurrent infections, and profoundly impaired oral wound healing. Through careful biochemistry and molecular approaches we have identified this child as having a novel defect in the scaffolding protein WDR1. Interestingly enough, we have found the same defect in a child seen her 25 years ago with similar pathology who and identified mutations in the same gene. These combined approaches continue to be productive and help us understand innate immunity and inflammation. These studies will help us understand several different infections, including fungal infections, at a molecular genetic and functional level.