Characterization of innate and IgE-mediated mast cell functions in honeybee venom allergy using Collaborative Cross mice

使用 Collaborative Cross 小鼠表征蜜蜂毒液过敏中的先天和 IgE 介导的肥大细胞功能

• 批准号: 10331200
• 负责人: Stephen Joseph Galli
• 金额: $ 55.18万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10331200
• 关键词: Accounting; Affinity; Allergens; Allergic; Allergic Disease; Allergic Reaction; Anaphylaxis; Animals; Antigens; Bacteria; Binding; Biological Models; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Candidate Disease Gene; Cell Degranulation; Cell physiology; Cell secretion; Cells; Cessation of life; Clinical; Coupled; Cytoplasmic Granules; Development; Disease; Dose; Exposure to; Food; Genes; Genetic; Genetic Variation; Growth Factor; Host Defense; Human; Hymenoptera; Hypersensitivity; IgE; IgE Receptors; IgG1; Immune; Immune response; Immunity; In Vitro; Individual; Inflammation; Inflammation Mediators; Insect Sting; Insecta; Life; Lipids; Measures; Mediating; Mediator of activation protein; Medicine; Modeling; Modification; Mus; Outcome; Parasites; Passive Cutaneous Anaphylaxis; Pathology; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Predisposition; Prevention approach; Production; Quantitative Trait Loci; Reaction; Regulatory Pathway; Research; Resistance; Resolution; Role; Severities; Shock; Signs and Symptoms; Snake Venoms; Staphylococcus aureus; Sting Injury; Strongyloides; System; Technology; Tissues; Toxic effect; Variant; Venoms; Work; Zebrafish; base; cardiovascular collapse; chemokine; confocal imaging; crosslink; cytokine; experience; experimental study; functional genomics; genetic analysis; genetic linkage analysis; genomic locus; improved; mast cell; novel; pathogenic bacteria; personalized approach; prevent; response; screening; trait

PROJECT SUMMARY/ABSTRACT We will use Collaborative Cross (CC) mice to define how genetic traits influence innate and/or IgE-mediated responses of mast cells (MCs) to honeybee venom. In IgE-dependent allergic reactions, crosslinking of MC high affinity IgE receptors (i.e., FceRI) by the binding of bivalent or multivalent allergen to antigen-specific IgE activates MCs to secrete three major classes of products: 1) preformed mediators stored in cytoplasmic granules, 2) newly synthesized lipid-derived mediators, and 3) cytokines, chemokines and growth factors. These MC products are responsible for many of the signs and symptoms of allergic diseases. MC activation, with or without the involvement of IgE, is also thought to contribute to the inflammation, tissue damage, and even fatal shock induced by envenomation. Components of hymenoptera venoms (e.g., honeybee venom), pharmaceutical agents, and foods are the most common triggers for anaphylaxis in humans. Many people have been sensitized to hymenoptera venoms and some unfortunate individuals react after exposure to such insect stings with serious systemic reactions and even fatal anaphylaxis. However, recent experiments in mice and zebra fish demonstrate that MC-derived proteases can degrade animal venoms and diminish their toxicity. Also, IgE/FceRI-mediated MC activation can enhance the survival of mice challenged with honeybee venom or a snake venom, or with S. aureus bacteria. Yet the benefits of “allergic immune responses”, mediated by IgE/MC- dependent mechanisms, have not been widely recognized. While the exact mechanisms determining whether the outcomes of hymenoptera envenomation are detrimental or favorable have been elusive, we know that genetic factors can significantly influence the development, progression, and severity of allergy and anaphylaxis. We hypothesize that genetic traits, by modulating the strength and/or composition of MC responses to honeybee venom and/or the Th2-IgE-MC immune axis, can influence the outcomes of honeybee stings. In this project, we propose to use genetically diverse CC mice to identify genetic modifiers regulating MC functions in insect venom allergy. In Aim 1, we will screen a panel of CC mice for their susceptibility to the toxicity of honeybee venom, development and features of venom-specific type 2 immunity, and induction of MC activation with or without crosslinking of venom-specific IgE/FceRI. We will also perform quantitative trait locus (QTL) mapping of the venom-induced phenotypes in CC mice to identify distinct genetic loci and novel regulators associated with MC-dependent susceptibility vs. resistance to honeybee venom. In Aim 2, we will confirm, in mouse MCs, important regulators of MC functions that are identified by screening CC mice using QTL analysis to assess the involvement of these regulators in innate and/or IgE-mediated MC functions. We think that the identification of genetic modifiers that distinguish beneficial vs. harmful effects of innate and/or allergic immune responses to honeybee venom will improve the treatment of severe reactions to insect venom and probably other disorders in which IgE and MCs have a critical role. 1

项目摘要/摘要 我们将使用协作杂交（CC）小鼠来定义遗传特征如何影响先天和/或IgE介导的 肥大细胞（MC）对蜜蜂毒液的反应。在IgE依赖性过敏反应中，MC的交联 高亲和力IgE受体（即FCERI）通过二价或多价过敏原与抗原特异性IgE结合 激活MC分泌三个主要产品：1）存储在细胞质中的介体 颗粒，2）新合成的脂质衍生的介质，以及3）细胞因子，趋化因子和生长因子。 这些MC产品负责过敏性疾病的许多体征和症状。 MC激活， 有或不参与IgE的情况下，也被认为会导致炎症，组织损伤和 甚至是由于衰老引起的致命休克。膜翅目毒液的成分（例如，蜜蜂毒液）， 药物和食物是人类过敏反应的最常见触发因素。很多人有 我们对膜翅目毒液敏感，一些不幸的人在暴露于这种昆虫后做出反应 具有严重的全身反应甚至致命过敏反应的刺痛。但是，最近在小鼠和 斑马鱼表明，MC衍生的蛋白酶可以降解动物毒液并降低其毒性。还， IgE/FCERI介导的MC激活可以增强honeybee毒液或A的挑战的小鼠的存活率 蛇毒，或与金黄色葡萄球菌细菌。然而，由IGE/MC-介导的“过敏免疫反应”的好处 依赖机制尚未得到广泛认可。而确切的机制决定了是否 膜翅目肌的结果是有害或有利的，难以捉摸，我们知道 遗传因素可以显着影响过敏的发展，进展和严重程度， 过敏反应。我们通过调节MC的强度和/或组成来假设遗传特征 对蜜蜂毒液和/或Th2-IgE-MC免疫轴的反应可能会影响 蜜蜂刺。在这个项目中，我们建议使用遗传学的CC小鼠识别遗传修饰剂 调节昆虫毒液过敏中的MC功能。在AIM 1中，我们将筛选一组CC小鼠的小鼠 对蜜蜂毒液的毒性的敏感性，毒液特异性2型免疫支持的发育和特征， 并在有或不使用毒液特异性IgE/FCERI的情况下诱导MC激活。我们也会表演 CC小鼠中毒液诱导的表型的定量性状基因座（QTL）映射，以识别不同的通用 基因座和新型调节剂与依赖MC的敏感性与对蜜蜂毒液的抗性相关。在 AIM 2，我们将在鼠标MC中确认MC函数的重要调节剂，这些调节剂是通过筛选CC识别的 使用QTL分析的小鼠评估这些调节因子与先天和/或IgE介导的MC的参与 功能。我们认为，鉴定区分有益和有害影响的遗传修饰符 对蜜蜂毒液的先天和/或过敏反应将改善对严重反应的治疗 昆虫毒液以及IgE和MC具有关键作用的其他疾病。 1