Clinical Studies Of Abnormal Host Defense

1)在本财年，由道格拉斯·库恩斯领导的我们的临床实验室-中性粒细胞监测实验室(NML)对56名疑似CGD患者进行了分子诊断研究。总共发现了29名新的X连锁CGD患者和11名常染色体CGD患者。通过对57名CGD患者或携带者的核酸测序确认了诊断。NML已经进行了1262次超氧化物检测和740次DHR检测，以支持与CGD相关的积极临床方案。 2)本课题组继续对新型革兰氏阴性CGD病原菌Granulibacter bethe densis进行临床和实验室研究。在2013财年，我们继续对CGD和其他疾病的MDH血清阳性进行筛查。我们已经确定了其他血清阳性的CGD患者，并正在评估恢复期血清阳性的进展情况。在与Joseph Fontana(NIH/NHLBI)的合作下，我们检测了结节病患者的颗粒状杆菌血清阳性。一项关于CGD患者和正常人的单核细胞和巨噬细胞对G.bethedensis的抗菌活性的研究已经发表(Chu等人，2013年，J免疫学)。 3)在2013财年期间，我们分析了美国国立卫生研究院#10-I-0029无创评估患有CGD和其他免疫系统疾病患者的动脉粥样硬化的数据(目前共有74名受试者)。动脉粥样硬化是心脏病的主要原因，被认为与心脏血管的炎症反应失调有关，并与过度产生活性氧物种(ROS)有关。我们假设，CGD患者，其吞噬细胞和其他细胞产生的活性氧物种不足，可能受到保护，不会发生动脉粥样硬化。这项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患者临床前动脉粥样硬化症状的发生率有显著差异，并提交了一份描述这些发现的手稿。 4)今年我们对CGD患者的B细胞功能进行了研究。我们在Matharu等人上报道了这一点。(2013，《临床免疫学》)，CGD患者和静脉注射脂多糖(LPS)的正常患者血清中B细胞激活因子(BAFF)浓度显著升高。这项研究还证明了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信息学开发实验室)合作进行了一项广泛的临床数据挖掘研究，以表征总免疫球蛋白水平和NIH临床中心CGD队列中的自身免疫球蛋白水平，并将这些参数与其他基因和表型数据相关联。进一步的分析还在进行中。 5)几年前，两名未确诊的疾病患者在NIH就诊，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年的时间范围内看到的另外2个患者家庭有相似之处。细胞研究显示，中性粒细胞形态异常，核突出，趋化性异常，细胞骨架结构异常。我们已经通过生化和分子方法鉴定了这种疾病的分子缺陷，并提交了这篇论文以供发表。

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.