Clinical Studies Of Abnormal Host Defense

1)在本财政年度，神经元监测实验室（NML），我们的临床实验室合作由道格拉斯Kuhns管理，已经对56例疑似CGD患者进行了分子诊断研究。 共发现29例新的X连锁CGD患者和11例常染色体CGD患者。 通过总共57名CGD患者或携带者的核酸测序证实了诊断。 NML已进行了1，262次超氧化物测定和740次DHR测定，以支持与CGD相关的有效临床方案。 2)我们的小组继续其临床和实验室研究的新革兰氏阴性CGD病原体，贝氏颗粒杆菌。在2013财年，我们继续筛查CGD和其他疾病的MDH血清阳性。 我们已经确定了其他血清阳性CGD患者，并正在评估恢复期血清阳性的进展。 与Joseph丰塔纳（NIH/NHLBI）合作，我们检测了结节病患者的颗粒杆菌血清阳性。 已经发表了来自CGD患者和正常受试者的单核细胞和巨噬细胞针对贝塞登革菌的抗微生物活性的研究（Chu等人，2013，J Immunology）。 3)在2013财年，我们分析了NIH方案#10-I-0029 CGD和其他免疫系统疾病患者动脉粥样硬化的无创评估（当前总计= 74例受试者）的数据。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。我们假设，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。 我们发现CGD患者动脉粥样硬化临床前体征的发生率存在显著差异，并提交了一份描述这些发现的手稿。 4)今年我们研究了CGD患者的B细胞功能。 我们在Matharu et al.（2013，Clinical Immunology）中报道了CGD患者血清中B细胞活化因子（BAFF）浓度的显著升高以及用脂多糖（LPS）静脉内治疗的正常患者。 本研究还证实了BAFF与急性期反应物的相关性。 由于BAFF在CGD中升高，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，并将这些参数与其他基因型和表型数据相关联。 尚待进一步分析。 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，并提交了这篇论文发表。

1) During this FY, the Neutrophil Monitoring Laboratory (NML), our clinical laboratory collaboration managed by Douglas Kuhns, has performed molecular diagnostic studies on 56 patients suspected of having CGD. A total of 29 new X-linked CGD patients were identified as well as 11 autosomal CGD patients. Diagnosis was confirmed by nucleic acid sequencing of a total of 57 CGD patients or carriers. The NML has performed 1,262 superoxide assays and 740 DHR assays to support active clinical protocols relating to CGD. 2) Our group continues its clinical and laboratory studies of the novel Gram-negative CGD pathogen, Granulibacter bethesdensis. During FY13, we continued screening for MDH seropositivity in CGD and other disorders. We have identified additional seropositive CGD patients and are evaluating the progression of seropositivity during convalescence. In collaboration with Joseph Fontana (NIH/NHLBI) we examined Granulibacter seropositivity in patients with sarcoidosis. A study of the antimicrobial activities of monocytes and macrophages from CGD patients and normal subjects against G.bethesdensis has been published (Chu et al., 2013, J Immunology). 3) During FY2013, we analyzed data from NIH Protocol #10-I-0029 Non-invasive Assessment of Atherosclerosis in Patients with CGD and other Disorders of the Immune System (current total = 74 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 hypothesized 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 was 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. We found significant differences in the incidences of pre-clinical signs of atherosclerosis in CGD patients and have submitted a manuscript describing these findings. 4) This year we studied B-cell function in CGD patients. We reported in Matharu et al. (2013, Clinical Immunology), a significant elevation in the concentrations of B-cell Activating Factor (BAFF) in the serum of CGD patients as well as in normal patients treated intravenously with lipopolysaccharide (LPS). This study also demonstrated the correlations of BAFF with acute phase reactants. Since BAFF is elevated in CGD, we collaborated with James Cimino (NIH Laboratory for Informatics Development) to perform an extensive clinical data-mining study to characterize both total and autoimmune immunoglobulin levels and in the NIH Clinical Center CGD cohort as well as correlate these parameters with other genotypic and phenotypic data. Further analysis is pending. 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 have submitted this paper for publication.