Clinical Studies Of Abnormal Host Defense

1)由道格拉斯库恩斯管理的我们的临床实验室合作组织神经元监测实验室（NML）通过蛋白质印迹法对38例受试者的分子缺陷进行了表征，并确定了29例新的CGD患者。 通过总共55名CGD患者或携带者的核酸测序证实了诊断。 2)本研究组参与了新型革兰氏阴性CGD病原体贝塞登颗粒杆菌的临床和实验室研究。在2012财年，我们已经纯化了患者血清识别的主要抗原甲醇脱氢酶（MDH），并已开始对该酶进行生物化学表征。与LCID合作，我们参与了CGD队列的血清学研究，使用免疫印迹和基于MDH的ELISA。这些研究已经确定了另外几名CGD患者，这些患者有可能既往感染过颗粒杆菌的证据，并为可能怀疑感染的其他患者人群的大规模筛查提供了工具。 该项目已发表在传染病杂志上。 3)在2012财年期间，我们在NIH方案#10-I-0029 CGD和其他免疫系统疾病患者的动脉粥样硬化无创评估中研究了另外24例患者（当前总计= 73例受试者）。动脉粥样硬化是心脏病的主要原因，被认为与心脏血管中的炎症失调有关，并且涉及活性氧（ROS）的过度产生。我们推测，CGD患者，他们的吞噬细胞和其他细胞的活性氧产生不足，可能是保护发展动脉粥样硬化。本研究的主要终点是通过CT、MRI和其他成像方法评估这些和其他先天性免疫功能障碍患者的动脉粥样硬化斑块形成/钙沉积。炎症性肠病提供了一组具有相似慢性炎症但具有完整ROS产生的患者用于比较。重要的是，这项研究可能首次在人类中确定ROS是否确实在动脉粥样硬化的发病机制中发挥作用，这一发现可能对一般人群具有广泛的重要性，因为它将为治疗干预提供经验证的分子靶点。 4)今年，我们开始了CGD中免疫球蛋白生产的研究。 最早报道的CGD患者的表型之一是观察到异常高量的IgG，并且IgG产生的失调已经被假设与这些受试者中自身免疫性疾病的频率增加有关。 我们与James Cimino（NIH信息学发展实验室）合作进行了一项广泛的临床数据挖掘研究，以表征NIH临床中心CGD队列中的总免疫球蛋白和自身免疫球蛋白水平以及发病率，并将这些参数与其他基因型和表型数据相关联。 在体外实验室研究测量B细胞功能和免疫球蛋白分泌揭示了潜在的重要差异之间的正常和CGD B细胞。 这项工作已提交出版。 5)几年前，在NIH发现了两名未确诊疾病的患者，其特征是感染增加和中性粒细胞减少。 这些受试者与LHD在30年的时间范围内观察到的其他2个患者家族相似。 细胞研究表明，异常的中性粒细胞形态与频繁的核突出，低于正常的趋化性，和异常的细胞骨架结构。我们已经确定了生物化学和分子的方法在这种疾病的分子缺陷，目前正在准备提交本出版物。

1) The Neutrophil Monitoring Laboratory (NML), our clinical laboratory collaboration managed by Douglas Kuhns, has characterized the molecular defects by western blotting of 38 subjects and identified 29 new CGD patients. Diagnosis was confirmed by nucleic acid sequencing of a total of 55 CGD patients or carriers. 2) Our group has been involved in the clinical and laboratory studies of the novel Gram-negative CGD pathogen, Granulibacter bethesdensis. During FY12, we have purified the dominant antigen recognized by patient sera, methanol dehydrogenase (MDH), and have begun the biochemical characterization of this enzyme. In collaboration with the LCID, we have participated in serologic studies of the CGD cohort using both immunoblotting and an ELISA based on MDH. These studies have identified several additional CGD patients with evidence of likely prior infections with Granulibacter and provide tools for larger-scale screening of other patient population in which this infection may be suspected. This project has been published in the Journal of Infectious Diseases. 3) During FY2012, we have studied 24 more patients on NIH Protocol #10-I-0029 Non-invasive Assessment of Atherosclerosis in Patients with CGD and other Disorders of the Immune System (current total = 73 subjects). Atherosclerosis, the major cause of heart disease, is thought to relate to dysregulated inflammation in the cardiac blood vessels and over production of reactive oxygen species (ROS) has been implicated. We hypothesize that CGD patients, who have deficient production of reactive oxygen species by their phagocytes and other cells, may be protected from developing atherosclerosis. The primary endpoint of this study is the assessment of atherosclerotic plaque formation/calcium deposition by CT, MRI and other imaging methodologies, in these and other patients with in-born disorders of immune function. Inflammatory bowel disease provides a group of patients with similar chronic inflammation but with intact ROS production for comparison. Importantly, this study may determine for the first time in humans, whether ROS indeed play a role in the pathogenesis of atherosclerosis, a finding that could have broad importance for the general population as it would provide a validated molecular target for therapeutic intervention. 4) This year we initiated a study of immunoglobulin production in CGD. One of the earliest reported phenotypes of CGD patients was the observation of abnormally high amounts of IgG and dysregulation of IgG production has been linked hypothetically to the increased frequency of autoimmune disorders in these subjects. We undertook, in collaboration with James Cimino (NIH Laboratory for Informatics Development), an extensive clinical data-mining study to characterize both total and autoimmune immunoglobulin levels and incidence in the NIH Clinical Center CGD cohort as well as correlate these parameters with other genotypic and phenotypic data. In vitro laboratory studies measuring B-cell function and immunoglobulin secretion have revealed potentially important differences between normal and CGD B-cells. This work has been submitted for publication. 5) Several years ago, two patients with an undiagnosed disease characterized by increased infections and neutropenia were seen at the NIH. These subjects bore similarities to 2 other families of patients seen over a time frame of 30 years by the LHD. Cellular studies indicated an abnormal neutrophil morphology with frequent nuclear herniations, subnormal chemotaxis, and aberrant cytoskeletal structure. We have identified by biochemical and molecular approaches the molecular defect in this disease and are currently preparing this publication for submission.