Epithelial Cell-dependent Activation of Human Basophils

人类嗜碱性粒细胞的上皮细胞依赖性激活

• 批准号: 9179854
• 负责人: JOHN T. SCHROEDER
• 金额: $ 24.3万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9179854
• 关键词: A549; Address; Affect; Affinity; Agonist; Allergens; Allergic; Allergic Disease; Antibodies; Antigens; Asthma; Atopic Dermatitis; Basophils; Binding; Biological Assay; Cell Communication; Cell Line; Cells; Cellular Structures; Child; Coculture Techniques; Connective Tissue Cells; Data; Dependency; Disease susceptibility; Epithelial Cells; Epithelium; Fibroblasts; Food; Galectin 3; Histamine; Histamine Release; Human; Hypersensitivity; IgE; IgE Receptors; Immune; Immune response; Inflammation; Inflammation Mediators; Interleukin-13; Interleukin-3; Interleukin-4; Kinetics; Laboratories; Learning; Leukocytes; Leukotriene C4; Link; Lung; Mediating; Mediator of activation protein; Mus; Nature; Organ; Pathogenesis; Play; Population; Probability; Production; Proteins; Reporting; Role; Signal Transduction; Skin; Small Interfering RNA; Solid; Source; Stimulus; Surface; Symptoms; TSLP gene; Testing; Transfection; Translating; United States; Urticaria; Virus; Work; anti-IgE; cell type; chemical release; clinically relevant; crosslink; cytokine; experience; granulocyte; in vivo; innovation; insight; knock-down; mouse model; novel; omalizumab; research study; response; skin disorder; stem; sugar

ABSTRACT Allergic diseases are caused by inappropriate immune responses to otherwise harmless antigens, and, in one form or another, affect up to 25% of the population in the United States, with a disproportionate increase especially among children with asthma and food-related hypersensitivities. While many immune cell types are implicated in the pathogenesis of allergic disease there is growing evidence that basophil granulocytes play a far greater role than initially thought. Indeed, a good portion of this excitement stems from work done ~20 years ago (including our own) showing that human basophils are a prolific source of IL-4 & IL-13 –cytokines central to the allergic diathesis. More recently, these findings have been confirmed and extended in vivo using mouse models. Moreover, some mouse models now show evidence of an “axis” whereby epithelial cell (EC)-derived cytokines (e.g. TSLP, but also IL-33 and IL-25) activate basophils to produce IL-4 (& IL-13) that is seemingly critical for initiating IgE synthesis and thus sensitization. In contrast, confirmatory results identifying TSLP as an activator of human basophils have not been forthcoming. In exploring this translational discrepancy, we have now uncovered a remarkably robust, and potentially unique, mode of EC- dependent activation of human basophils that occurs independently of known EC-derived cytokines. Basophils co-cultured with EC of lung origin (A549 cells) in the presence of IL-3 secrete high IL-4/IL- 13 levels yet to be achieved with other modes of stimulation. This response is preceded by mediator (histamine) release, but with delayed kinetics compared to that seen with standard FcεRI-dependent activation. New evidence points to an IgE-binding protein on EC (e.g. galectin-3) that is mediating this unique mode of basophil activation. Two aims are proposed to further characterize this novel response. Aim 1 explores the nature of this interaction, hypothesizing the need for cell-to-cell interactions. The role of IL-3 priming of basophils and the requirement for IgE are also to be investigated. We proposed experiments also testing whether EC activate other IgE-bearing cells (e.g. mast cells) for similar activity. And, other EC lines (including normal EC) are to be investigated for mediating basophil activation, hypothesizing that “danger signals” (viruses, cytokines, TLR agonists) play an important role in enabling the latter to possess a similar capacity. Aim 2 studies are to explore the role of known IgE-binding proteins such as galectin-3 (but also CD23b, and potentially novel ones), in meditating this activity. Overall, our unique experience working with human basophils makes this R21 application highly innovative, and increases the probability that novel, significant, and clinically relevant findings will be discovered.

摘要 过敏性疾病是由不适当的免疫反应引起的， 抗原，并且以一种或另一种形式影响美国高达25%的人口， 特别是在患有哮喘和与食物有关的过敏症的儿童中。 虽然许多免疫细胞类型与过敏性疾病的发病机制有关，但越来越多的免疫细胞类型被发现与过敏性疾病有关。 嗜碱性粒细胞发挥的作用比最初认为的要大得多。的确， 这种兴奋的一部分源于20年前（包括我们自己）所做的工作， 人嗜碱性粒细胞是IL-4和IL-13 -细胞因子的丰富来源，这些细胞因子对变应性素质至关重要。更 最近，这些发现已被证实，并在体内扩大使用小鼠模型。此外，委员会认为， 一些小鼠模型现在显示出“轴”的证据， (e.g. TSLP，还有IL-33和IL-25）激活嗜碱性粒细胞产生IL-4（和IL-13），这似乎是 对于启动IgE合成并因此致敏至关重要。相比之下，确认结果表明， TSLP作为人类嗜碱性粒细胞的激活剂尚未出现。在探索这种翻译 差异，我们现在已经发现了一个非常强大的，潜在的独特的，EC模式- 人嗜碱性粒细胞的依赖性活化，其独立于已知的EC衍生的细胞因子而发生。 在IL-3存在下与肺源EC（A549细胞）共培养的嗜碱性粒细胞分泌高IL-4/IL- 13个水平尚未达到与其他模式的刺激。此响应之前是调解人 （组胺）释放，但与标准Fcε RI依赖性 activation.新的证据指出EC上的IgE结合蛋白（例如半乳糖凝集素-3）介导了这一过程。 嗜碱性粒细胞激活的独特模式。两个目标提出进一步表征这部小说 反应目的1探讨了这种相互作用的性质，假设需要细胞对细胞的相互作用。 交互.嗜碱性粒细胞的IL-3引发作用和对IgE的需求也有待进一步研究。 研究了我们提出的实验也测试EC是否激活其他携带IgE的细胞（例如， 肥大细胞）的相似活性。并且，将调查其他EC线（包括正常EC）， 介导嗜碱性粒细胞活化，假设“危险信号”（病毒、细胞因子、TLR激动剂） 在使后者拥有类似能力方面发挥重要作用。目的2研究旨在探索 已知的IgE结合蛋白如半乳糖凝集素-3（还有CD 23 b，以及潜在的新蛋白）的作用， 冥想这个活动。总的来说，我们与人类嗜碱性粒细胞合作的独特经验使得 R21应用高度创新，并增加了新的，重要的，临床上 将发现相关发现。