Laboratory Studies of Abnormal Host Defense and Immunoregulatory Diseases

宿主防御异常和免疫调节疾病的实验室研究

• 批准号: 8745571
• 负责人: Kol Zarember
• 金额: $ 11.37万
• 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/8745571
• 关键词: Anti-Inflammatory Agents; Anti-inflammatory; Antimicrobial Cationic Peptides; Biochemical; Carbohydrates; Cells; Chronic Granulomatous Disease; Collaborations; Complement; Complex; Diagnostic; Disease; Enzymes; Gel Chromatography; Goals; Host Defense; Human; Immune; Immune system; Immunity; Infection; Inflammatory; Interferons; Ion Exchange; Laboratory Study; Leukocytes; Lipopolysaccharides; Methanol; Modeling; Molecular Models; NADPH Oxidase; National Institute of Diabetes and Digestive and Kidney Diseases; Normal Cell; Organism; Oxidoreductase; Paper; Pathogenesis; Patients; Publications; Research; Residual state; Resistance; Serologic tests; Structure; Substrate Specificity; Testing; United States National Institutes of Health; Universities; congenital immunodeficiency; cytokine; inhibitor/antagonist; innate immune function; killings; macrophage; microbial; microorganism; molecular modeling; monocyte; neutrophil; novel; pathogen

1) To study the determinants of monocyte and macrophage host defenses against infection by the novel bacterial pathogen, Granulibacter bethesdensis Our recent publication (Zarember et al., Infection and Immunity, 2012) demonstrated that killing of G. bethesdensis by neutrophils from patients with chronic granulomatous disease (CGD) was defective and that this Gram-negative pathogen was remarkably resistant to complement and to cationic antimicrobial peptides. During FY13, Jessica Chu (Postdoctoral IRTA) completed a study demonstrating that CGD human monocytes and monocyte-derived macrophages were unable to kill G. bethesdensis normally and that this organism could persist in CGD macrophages. This paper also demonstrated a positive correlation between microbial killing by IFNgamma-treated monocytes and residual neutrophil NADPH oxidase activity. This finding has potential diagnostic implications in the use of IFN (Actimune) to treat CGD patients. 2) G. bethesdensis Lipopolysaccharide (LPS) During our studies of the interaction of Granulibacter bethesdensis with immune cells, we found that this organism is remarkably hypostimulatory of the human innate immune system, both in terms of weak activation of the NADPH oxidase and poor stimulation of cytokine secretion. We are collaborating with Yossi Shiloach (NIDDK) and Russell Carlson of the University of Georgia Complex Carbohydrate Research Center to complete the purification and structural characterization of the atypical lipopolysaccharide (LPS) of this organism and determine whether it acts as an anti-inflammatory inhibitory LPS. 3) G.bethesdensis Methanol Dehydrogenase In order to develop our serological testing for G.bethesdensis infection (see ZIA AI000155-36), we purified Methanol Dehydrogenase from this organism. Using ion exchange and gel filtration, highly enriched enzyme was prepared and biochemical testing is underway to identify substrate specificity, identify inhibitors, and s indicate a much broader substrate specificity that originally thought. In collaboration with Peter Steinbach (NIH Center for Molecular Modeling), we have modeled the structure of G. bethesdensis MDH.

1) 研究单核细胞和巨噬细胞宿主防御新型细菌病原体贝塞斯登细粒杆菌感染的决定因素 我们最近发表的文章（Zarember 等人，感染与免疫，2012）证明，慢性肉芽肿病 (CGD) 患者的中性粒细胞对贝塞斯登氏菌的杀灭作用是有缺陷的，并且这种革兰氏阴性病原体对补体和阳离子抗菌肽具有显着的抗性。 2013 财年，Jessica Chu（IRTA 博士后）完成了一项研究，证明 CGD 人类单核细胞和单核细胞衍生的巨噬细胞无法正常杀死 G. bethesdensis，并且该生物体可以在 CGD 巨噬细胞中持续存在。 该论文还证明了 IFNγ 处理的单核细胞杀灭微生物与残留中性粒细胞 NADPH 氧化酶活性之间的正相关性。 这一发现对于使用 IFN (Actimune) 治疗 CGD 患者具有潜在的诊断意义。 2) G. bethesdensis 脂多糖 (LPS) 在我们对 Bethesdensis 颗粒杆菌与免疫细胞相互作用的研究中，我们发现这种生物体对人类先天免疫系统的刺激作用显着不足，无论是 NADPH 氧化酶的激活较弱，还是细胞因子分泌的刺激较差。我们正在与佐治亚大学复合碳水化合物研究中心的 Yossi Shiloach (NIDDK) 和 Russell Carlson 合作，完成该生物体非典型脂多糖 (LPS) 的纯化和结构表征，并确定其是否具有抗炎抑制 LPS 的作用。 3) G.bethesdensis 甲醇脱氢酶 为了开发 G.bethesdensis 感染的血清学检测（参见 ZIA AI000155-36），我们从该生物体中纯化了甲醇脱氢酶。 使用离子交换和凝胶过滤，制备了高度富集的酶，并且正在进行生化测试，以鉴定底物特异性、鉴定抑制剂，并且表明比最初认为的更广泛的底物特异性。我们与 Peter Steinbach（NIH 分子建模中心）合作，对 G. bethesdensis MDH 的结构进行了建模。