Eosinophils and Eosinophil Activation in the Pathogenesis of Human Disease

人类疾病发病机制中的嗜酸性粒细胞和嗜酸性粒细胞激活

基本信息

批准号：
8946502
负责人：
Amy Klion
金额：
$ 104.48万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
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/8946502
关键词：
Adrenal gland hypofunction Adverse effects Aftercare Amyotrophic Lateral Sclerosis Angioneurotic Edema Antibodies Ascaridil Asthma Benign Biological Biological Markers Blood Bone Marrow Cells CD3 Antigens CD34 gene CD4 Positive T Lymphocytes Cells Cessation of life Childhood Clinical Clinical Treatment Clinical Trials Clinical Trials Design Comparative Study DNA Sequence Rearrangement Data Analyses Diethylcarbamazine Disease Disease remission Disseminated eosinophilic collagen disease Dose Double-Blind Method Drug Hypersensitivity Enrollment Eosinophil Granule Proteins Eosinophilia Eosinophilic leukemia Etiology Experimental Autoimmune Encephalomyelitis Extrinsic asthma Family Glucocorticoids Goals Helminths Hepatitis Homeostasis Human Hypersensitivity Imatinib Immunologics In Vitro In complete remission Infection Interleukin-5 International Ivermectin Liquid substance Loa Loa loa Loiasis Lymphocyte Manuscripts Measures Mediating Methods Microfilaria Monitor Monkeys Myeloproliferative disease Natural Killer Cells Onchocerciasis PDGFRA gene Parasitic infection Pathogenesis Patients Pattern Periodicity Pharmaceutical Preparations Phase III Clinical Trials Plasma Population Rare Diseases Reaction Refractory Role Serum Severe Adverse Event Societies Steroid Resistance Subgroup Surface Suspension substance Suspensions Symptoms Syndrome System T-Lymphocyte TNFSF5 gene Therapeutic Tissues United States National Institutes of Health Work chemokine clinical efficacy cohort cytokine double-blind placebo controlled trial effective therapy eosinophil human SIGLEC8 protein human disease in vivo inhibitory surface receptor killings mRNA Expression mast cell mastocytosis meetings mepolizumab neoplastic neutrophil novel novel diagnostics open label peripheral blood placebo controlled study pre-clinical preclinical study prevent receptor response stem tool

项目摘要

Using a cohort of more than 350 subjects with a wide variety of eosinophilic disorders, ranging from benign eosinophilia to eosinophilic leukemia, we have continued to identify and characterize novel subgroups of patients with eosinophilia. Episodic Angioedema with Eosinophilia (EAE; Gleichs Syndrome) is a rare disorder characterized by episodes of angioedema and eosinophilia that occur at monthly intervals and resolve spontaneously without therapy. To characterize the involvement of lineages other than eosinophils in the etiology and pathogenesis of EAE, nine subjects with suspected EAE were evaluated at the NIH. Cycling of multiple lineages, including neutrophils, lymphocytes and eosinophils, was observed in 5 subjects with EAE with a periodicity of 25-35 days. An aberrant CD3-CD4+ T cell population was detected in all 5 subjects, and TCR rearrangement studies showed a clonal pattern in 3 subjects. A peak of type II cytokines was detected in the serum of EAE subjects prior to onset of symptoms and eosinophil cycling and corresponded to ex-vivo type II cytokines detected intracellularly in CD3+CD4+CD154+ T cells. Although the etiology of EAE is not yet known, multiple lineages, including lymphocytes, neutrophils and mast cells, are involved and may be related to disease pathogenesis. Whether these cells act directly or promote eosinophilia and eosinophil activation remains to be elucidated. Other subgroups of eosinophilic subjects currently under study include families with autosomal dominant eosinophilia, pediatric patients, and a cohort of subjects with eosinophilic hepatitis. Therapy for eosinophilic disorders remains a primary focus of our group. In the past year, we have initiated two new clinical trials for the treatment of HES. The first of these is a placebo-controlled, double-blind trial of benralizumab (MedImmune/Astra Zeneca), an afucosylated antibody to IL-5 receptor alpha that has shown clinical efficacy in patients with eosinophilic asthma. The current trial stems, in part, from our prior pre-clinical work examining IL-5 receptor levels in patients with eosinophilia and/or mastocytosis (Wilson et al. J Allergy Clin Immunol 2011). To date 6 of 20 planned subjects have been enrolled. The second trial is an open-label trial of dexpramipexole (Knopp Biosciences) in patients with steroid-resistant HES. Dexpramipexole was developed for the treatment of amyotrophic lateral sclerosis (ALS). Although it was well tolerated, dexpramipexole failed to show efficacy in a large phase 3 trial for this disease. It did, however, appear to selectively lower blood eosinophils, which prompted the current proof-of-concept trial in HES. To date, 4 of the 10 planned subjects have been enrolled. In addition to the clinical trials described above, we have analyzed data from an ongoing clinical trial of imatinib in subjects with FIP1L1/PDGFRA-positive myeloproliferative neoplasm (FP), PDGFRA-negative HES with myeloproliferative features (MP) and steroid-resistant without myeloproliferative features (SR) (Khoury et al. presented at the annual meeting of the International Eosinophil Society; manuscript submitted). Overall, imatinib response rates were 100% in the FP group (n= 16), 50% in the MP group (n=13) and 0% in the SR group (n=16). The presence of ≥ 4 myeloproliferative features was the sole predictor of response. After ≥ 18 months in complete remission, imatinib was tapered and discontinued in 7 FP and 1 MP subjects. Five subjects remain in remission off therapy for a median of 13 months (range 2-21 months). We have also continued to enroll subjects in a double-blind clinical trial designed to assess the utility of a glucocorticoid challenge in determination of glucocorticoid responsiveness in HES and in a clinical trial of mepolizumab (humanized anti-IL-5 antibody) for treatment-refractory HES. We have also begun to explore the therapeutic potential of several new antibodies with the potential to target eosinophils, including antibodies to the eosinophil specific surface receptor, EMR1 (Legrand J et al. Allergy Clin Immunol, 2014), and the inhibitory surface receptor, siglec-8. In a pre-clinical study, analysis of blood and bone marrow cells from healthy and eosinophilic donors and in vitro-differentiated CD34(+) cells confirmed restriction of human EMR1 surface and mRNA expression to mature eosinophils. Tissue eosinophils also expressed EMR1. Although EMR1 was highly expressed on eosinophils from all subjects, surface expression was negatively correlated with absolute eosinophil counts (r = -0.46, P < .001), and soluble plasma levels correlated positively with absolute eosinophil counts (r = 0.69, P < .001), suggesting modulation of EMR1 in vivo. Nevertheless, afucosylated anti-EMR1 mAb (Kalobios, Inc.) dramatically enhanced natural killer cell-mediated killing of eosinophils from healthy and eosinophilic donors and induced a rapid and sustained depletion of eosinophils in monkeys. Helminth infections can associated with dramatic eosinophilia and complications that are indistinguishable from HES. This is particularly true in loiasis where symptoms are associated with increased eosinophil granule protein levels (indicative of eosinophil activation and degranulation) as well as elevated levels of eosinophil-associated cytokines (Herrick et al. Clin Inf Dis, in press). In some infections, eosinophilia and eosinophil-related symptoms can be exacerbated by effective treatment. For example, treatment of the filarial infection loiasis with diethylcarbamazine (DEC) can be associated with severe adverse events, including death, that are accompanied by a dramatic rise in eosinophilia and are positively correlated with the number of circulating microfilariae in the blood. In the past year, we have continued our double-blind, placebo-controlled study of the anti-IL-5 antibody, reslizumab, to prevent post-treatment side effects of DEC in patients with Loa loa infection. Because similar side effects have been seen in Loa infected patients during mass administration of ivermectin for onchocerciasis control, we also conducted a comparative study of post-treatment reactions following single dose ivermectin or single dose DEC. Although clinical symptoms were similar between the two groups post-treatment, there was a significant difference in peak percentage eosinophil change (GM change + 1.865 in DEC group versus + 2.572 in IVM group, p <.05) and the pattern of post-treatment neutrophil count between the two groups. The microfilarial counts decreased dramatically in both groups following therapy, but mf were cleared more effectively in the DEC group. Cytokine and chemokine profiles also suggest differences in the immunologic responses following the two drugs. Finally, in the past year, we have continued to look for biomarkers of eosinophilic disease activity for use in monitoring responses to therapy and helping to dissect out the mechanisms of eosinophil pathogenesis. To this end, a multiplex suspension array system was developed to simultaneously measure the concentrations of 4 eosinophil granule proteins (MBP, ECP, EDN and EPO) in serum and other biological fluids and culture supernatants (Makiya et al. J Immunol Methods 2014).

我们使用350多名患者的队列，这些受试者患有各种嗜酸性疾病，从良性嗜酸性粒细胞到嗜酸性白血病，我们继续识别和表征嗜酸性粒细胞菌患者的新型亚组。嗜酸性粒细胞（EAE; Gleichs综合征）的发作性血管性水肿是一种罕见的疾病，其特征是血管性水肿和嗜酸性粒细胞增生的发作，每月以每月的间隔发生并在没有治疗的情况下自发解决。为了表征除嗜酸性粒细胞以外的其他谱系在EAE的病因和发病机理中的参与，在NIH上评估了九个具有可疑EAE的受试者。在5名EAE中观察到多个谱系的循环，包括中性粒细胞，淋巴细胞和嗜酸性粒细胞，周期性为25-35天。在所有5名受试者中都检测到异常的CD3-CD4+ T细胞群，TCR重排研究显示3名受试者的克隆模式。在症状和嗜酸性粒细胞循环发作之前，在EAE受试者的血清中检测到II型细胞因子的峰值，并对应于CD3+CD4+CD154+T细胞中细胞内检测到的Ex-Vivo II型细胞因子。尽管尚不清楚EAE的病因，但包括淋巴细胞，中性粒细胞和肥大细胞在内的多个谱系涉及，并且可能与疾病的发病机理有关。这些细胞是直接起作用还是促进嗜酸性粒细胞和嗜酸性粒细胞活化尚待阐明。目前正在研究的其他嗜酸性粒细胞受试者的其他亚组包括常染色体显性嗜酸性粒细胞，小儿患者和一系列患有嗜酸性肝炎的受试者。嗜酸性疾病的治疗仍然是我们小组的主要重点。在过去的一年中，我们启动了两项新的临床试验以治疗HES。其中第一个是安慰剂对照的双盲试验（Medimmune/astra Zeneca），这是一种抗IL-5受体α的抗糖基化抗体，在嗜酸性粒细胞性哮喘患者中显示出临床疗效。当前的试验部分源于我们先前的临床前研究，研究了嗜酸性粒细胞增多症患者的IL-5受体水平（Wilson等人Jallergy Clin Immunol 2011）。截至迄今为止，已招募了20名计划的主题中的6个。第二次试验是对耐药HES患者的右甲己烯（KNOPP Biosciences）的开放标签试验。开发了右氨己烯，用于治疗肌萎缩性侧索硬化症（ALS）。尽管耐受性良好，但在该疾病的大型3期试验中，右甲苯二曲未能显示出疗效。但是，它确实有选择地降低了血液嗜酸性粒细胞，这促使当前的概念证明试验在HES中。迄今为止，已经招募了10个计划的主题中的4个。 In addition to the clinical trials described above, we have analyzed data from an ongoing clinical trial of imatinib in subjects with FIP1L1/PDGFRA-positive myeloproliferative neoplasm (FP), PDGFRA-negative HES with myeloproliferative features (MP) and steroid-resistant without myeloproliferative features (SR) (Khoury et al. presented at the国际嗜酸性粒细胞协会的年度会议；总体而言，FP组的伊马替尼反应率为100％（n = 16），MP组为50％（n = 13），在SR组中为0％（n = 16）。 ≥4个骨髓增生特征的存在是响应的唯一预测指标。完全缓解≥18个月后，伊马替尼在7 FP和1 MP受试者中逐渐减少并停用。中位13个月（2-21个月）仍处于缓解治疗中的五名受试者。我们还继续将受试者参与了一项双盲临床试验，旨在评估糖皮质激素挑战在确定HES和Mepolizumab（人源化抗IL-5抗体）的临床试验中的糖皮质激素反应性方面的实用性。我们还开始探索几种新抗体的治疗潜力，具有靶向嗜酸性粒细胞的潜力，包括嗜酸性粒细胞特异性表面受体EMR1的抗体（Legrand J等人Allergy Clin Immunol，2014），以及抑制性表面受体Siglec-8。在一项临床前研究中，对健康和嗜酸性供体的血液和骨髓细胞的分析以及体外分化的CD34（+）细胞证实了人EMR1表面和mRNA表达对成熟嗜酸性粒细胞的限制。组织嗜酸性粒细胞也表示EMR1。尽管EMR1在所有受试者的嗜酸性粒细胞上都高度表达，但表面表达与绝对嗜酸性粒细胞计数呈负相关（r = -0.46，p <.001），并且可溶性血浆水平与绝对嗜酸性粒细胞计数呈正相关（r = 0.69，p <.001），建议模块化emr1 ins in emr1 ins in viv。然而，抗糖基化的抗EMR1 MAB（Kalobios，Inc。）急剧增强了自然杀手细胞介导的嗜酸性粒细胞的杀死，从健康和嗜酸性粒细胞供体中杀死嗜酸性粒细胞，并引起猴子中嗜酸性粒细胞的快速且持续的耗竭。蠕虫感染可能与剧烈的嗜酸性粒细胞和并发症有关，与HES无法区分。尤其是在虫病中，症状与嗜酸性粒细胞颗粒蛋白水平升高（指示嗜酸性粒细胞激活和脱粒）以及与嗜酸性粒细胞相关的细胞因子的水平升高有关（Herrick等人的Clin Inf dis，in Press）。在某些感染中，嗜酸性粒细胞和嗜酸性粒细胞相关的症状可以通过有效治疗加剧。例如，用二乙基钙化（DEC）处理丝状感染状态性（DEC）可能与严重的不良事件（包括死亡）相关，这些事件伴随着嗜酸性粒细胞增多症的急剧上升，并且与血液中循环的微部门循环的微部门数量呈正相关。在过去的一年中，我们继续对抗IL-5抗体Reslizumab进行双盲，安慰剂对照研究，以防止DEC对LOA LOA感染患者的DEC后处理后副作用。由于在大量给药伊维菌素对控制的伊维菌素期间，在LOA感染的患者中也有类似的副作用，因此我们还对单剂量伊维菌素或单剂量DEC进行了对治疗后反应的比较研究。尽管两组处理后两组之间的临床症状相似，但峰值百分比嗜酸性粒细胞变化有显着差异（DEC组的GM变化 + 1.865与IVM组中的 + 2.572，P <.05）和两组之间中性粒细胞计数的模式。在治疗后两组中，微虫计数均大幅下降，但在DEC组中更有效地清除了MF。细胞因子和趋化因子谱也表明两种药物后的免疫反应差异。最后，在过去的一年中，我们继续寻找嗜酸性疾病活性的生物标志物，用于监测对治疗的反应并帮助剖析嗜酸性粒细胞发病机理的机制。为此，开发了一个多重悬浮阵列系统，以同时测量血清和其他生物液和培养上清液中4种嗜酸性粒细胞颗粒蛋白（MBP，ECP，EDN和EPO）的浓度（Makiya等人，J Immunol方法2014）。