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: 10681390
• 负责人: Stephen Joseph Galli
• 金额: $ 52.46万
• 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/10681390
• 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; Maps; Measures; Mediating; Mediator; Medicine; Modeling; Modification; Mus; Outcome; Parasites; Passive Cutaneous Anaphylaxis; Pathology; Peptide Hydrolases; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Predisposition; Prevention approach; Probability; Production; Proliferating; Quantitative Trait Loci; Reaction; Regulatory Pathway; Research; Resistance; Resolution; Role; Severities; Shock; Signs and Symptoms; Snake Venoms; Staphylococcus aureus; Sting Injury; Strongyloides; System; Tissues; Toxic effect; Variant; Venoms; Work; Zebrafish; 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; prevent; response; screening; technology platform; 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

项目概要/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 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 分泌三类主要产物：1）储存在细胞质中的预制介质 颗粒，2) 新合成的脂质衍生介质，以及 3) 细胞因子、趋化因子和生长因子。 这些 MC 产品会导致过敏性疾病的许多症状和体征。 MC激活， 无论有或没有 IgE 的参与，也被认为会导致炎症、组织损伤和 甚至因中毒而引起致命的休克。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最常见的诱因。很多人都有 对膜翅目毒液敏感，一些不幸的个体在接触这种昆虫后会发生反应 蜇伤会引起严重的全身反应，甚至致命的过敏反应。然而，最近在老鼠身上进行的实验 斑马鱼证明，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 激活可以提高受到蜜蜂毒液或 蛇毒或金黄色葡萄球菌。然而，由 IgE/MC 介导的“过敏性免疫反应”的好处- 依赖机制尚未得到广泛认可。虽然确切的机制决定是否 膜翅目毒液的结果是有害还是有利一直难以捉摸，我们知道 遗传因素可以显着影响过敏的发生、进展和严重程度 过敏反应。我们假设遗传特征通过调节 MC 的强度和/或组成 对蜜蜂毒液和/或 Th2-IgE-MC 免疫轴的反应，可以影响以下结果： 蜜蜂蜇人。在这个项目中，我们建议使用基因多样化的 CC 小鼠来识别基因修饰剂 调节 MC 在昆虫毒液过敏中的功能。在目标 1 中，我们将筛选一组 CC 小鼠 对蜜蜂毒液毒性的易感性、毒液特异性2型免疫的发展和特征， 以及在有或没有毒液特异性 IgE/FceRI 交联的情况下诱导 MC 激活。我们还将表演 CC 小鼠毒液诱导表型的数量性状位点 (QTL) 作图，以鉴定不同的遗传 与 MC 依赖性易感性与蜜蜂毒液抗性相关的基因座和新型调节因子。在 目标 2，我们将在小鼠 MC 中确认通过筛选 CC 鉴定出的 MC 功能的重要调节因子 小鼠使用 QTL 分析来评估这些调节因子在先天和/或 IgE 介导的 MC 中的参与 功能。我们认为，识别出区分有益与有害影响的基因修饰剂 对蜜蜂毒液的先天和/或过敏性免疫反应将改善对蜜蜂毒液的严重反应的治疗 昆虫毒液以及 IgE 和 MC 可能发挥关键作用的其他疾病。 1

项目成果

Conditional neutrophil depletion challenges their contribution to mouse models of anaphylaxis.

条件性中性粒细胞耗竭挑战了它们对小鼠过敏反应模型的贡献。

• DOI: 10.1111/all.15738
• 发表时间: 2023
• 期刊: Allergy
• 影响因子: 12.4
• 作者: Stackowicz,Julien;Gillis,CaitlinM;Godon,Ophélie;Iannascoli,Bruno;Conde,Eva;Leveque,Edouard;Worrall,WilliamPM;Galli,StephenJ;Bruhns,Pierre;Reber,LaurentL;Jönsson,Friederike
• 通讯作者: Jönsson,Friederike