Rapid, safe suppression of IgE-mediated disease with monovalent anti-ceRIa mAb

使用单价抗 ceRIa mAb 快速、安全地抑制 IgE 介导的疾病

• 批准号: 10645062
• 负责人: FRED Douglass FINKELMAN
• 金额: $ 54.12万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-05 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10645062
• 关键词: Adverse effects; Affect; Affinity; Allergens; Allergic; Allergic Disease; Anaphylaxis; Animal Model; Animals; Antibodies; Antibody-mediated protection; Antigens; Basophils; Binding; Blood Cells; Cells; Clinical; Clinical Trials; Complement; Complement Receptor; Cross Reactions; Disease; Dissociation; Dose; Endocytosis; Excision; Exposure to; Flow Cytometry; Food Hypersensitivity; Granulocyte-Macrophage Colony-Stimulating Factor; Hour; Human; Hypersensitivity; IgE; IgE Receptors; IgG1; Immunodeficient Mouse; Immunoglobulin Class Switching; Immunoglobulin Constant Region; Immunoglobulin G; In Vitro; Injections; Interleukin 4 Receptor; Life; Macaca mulatta; Mediating; Modeling; Monoclonal Antibodies; Morbidity - disease rate; Mus; Persons; Preparation; Prevalence; Procedures; Process; Production; Protein Tyrosine Kinase; Protocols documentation; Reaction; Reagent; Safety; Serum; Signal Transduction; Stem Cell Factor; Surface; Testing; Transgenic Mice; Transgenic Organisms; Umbilical Cord Blood; Variant; Work; anergy; crosslink; cytokine; desensitization; humanized mouse; improved; in vivo; mast cell; mouse model; mutant; passive sensitization; prevent; receptor; reconstitution

This proposal tests the hypothesis that sustained, limited in vivo crosslinking of the high affinity IgE receptor, FcεRI, on mast cells (MCs) and basophils (BAs) rapidly desensitizes these cells and safely and effectively prevents IgE-mediated diseases, such as food allergy and anaphylaxis. Rapid desensitization (RD) is traditionally a procedure in which MCs and BAs from people who have IgE-mediated allergy to a specific allergen are made temporarily unresponsive to that allergen by exposure to increasing allergen concentrations over several hours. We modified RD by inoculating mice with increasing concentrations of an anti FcεRI α chain mAb. These studies showed that: (1) RD with anti-FcεRIα mAb is effective, safer and longer lasting than RD with allergen; (2) RD with anti-FcεRIα mAb is antigen-nonspecific; (3) MC unresponsiveness can be safely sustained by maintaining serum levels of this mAb; (4) this process works with anti-human (hu) FcεRIα mAbs in mice that express hu, rather than mouse FcεRIα (huFcεRIα mice); and (5) RD depends both on induction of MC anergy and removal of MC FcεRI and IgE. Recently, we have found that IgE-mediated anaphylaxis can be fully suppressed in huFcεRIα mice, without adverse effect, by repeatedly injecting <1 µg of IE7, a mouse mAb that binds to huFcεRIα regardless of FcεRI association with IgE. These mice can also be safely and dose- dependently desensitized by a single injection of a monovalent (mv) form of IE7 that has human IgG1-derived constant regions (mv huIgG1 IE7). mv huIgG1 IE7 also depletes blood BAs, and removes ~90% of MC IgE, but little FcεRIα. The loss of MC IgE and particularly, its disproportionate loss compared to FcεRI, was unexpected, because IE7 does not cause MCs to lose IgE in vitro and this mv Ab should only crosslink FcεRI if the Ab becomes multivalent by aggregating or by binding to cellular Fcγ or complement receptors. This proposal builds on these observations. Aim 1 uses flow cytometry, structural studies, anti-FcγR mAb, and hu IgG isotype switch variants of mv IE7 to determine whether mv huIgG1 IE7 crosslinks FcεRI in vivo and, if so, through what mechanism(s). Aim 1 also evaluates whether mv huIgG1 IE7 aggregation or internalization of IgE-loaded FcεRI is required for the disproportionate in vivo loss of MC IgE. Aim 2 applies Aim 1 results to use mv IE7 to safely desensitize actively sensitized, hyperallergic huFcεRIα mice that express a mutant IL-4 receptor as well as passively sensitized hu cord blood-reconstituted, hu cytokine-secreting, immunodeficient mice that develop large numbers of activated hu MCs and BAs; both models have been difficult to desensitize with increasing doses of divalent IE7. Aim 3 applies aim 2 results to desensitize rhesus monkeys, which express FcεRIα that reacts with IE7. Together, these aims should optimize the use of mv anti- FcεRIα mAb for RD and suggest whether and how it could be used to suppress hu IgE-mediated disease.

该提案检验了以下假设，即高亲和力IgE的体内交联有限 受体FcεRI，在肥大细胞（MCS）和嗜碱性粒细胞（BAS）上迅速使这些细胞脱敏，安全和 快速脱敏（RD） 传统上是一个程序，其中MC和BAS来自对IgE介导的过敏的人对特定的程序 过敏原通过暴露于过敏原浓度而暂时对该过敏原无反应 多个小时。我们通过以抗FcεRIα浓度增加的小鼠接种小鼠来改变RD 连锁mab。这些研究表明：（1）抗抗FcεRIαmAb的rd有效，更安全且持久比 RD与过敏原； （2）抗FcεRIαmAb的RD是抗原非特异性的； （3）MC无反应性可以安全 通过保持该单元的血清水平来维持； （4）此过程与抗人（HU）FCεRIαmAb一起使用 在表达HU的小鼠中，而不是小鼠FcεRIα（HUFCεRIα小鼠）； （5）RD都取决于诱导 MC Anergy和MCFCεRI和IgE的去除。最近，我们发现IgE介导的过敏反应可能是 通过反复注射<1 µg的IE7，小鼠mAb，完全抑制了HUFCεRIα小鼠，没有不利影响 与HUFCεRIα结合，无论FcεRI与IgE的关联如何。这些小鼠也可以安全和剂量 - 通过单次注入具有人IgG1衍生的IE7的单价（MV）形式的脱敏 恒定区域（MV Huigg1 IE7）。 MV Huigg1 IE7也会耗尽血液，并去除约90％的MC IgE，但 少量FcεRIα。 MC IgE的损失，尤其是其与FCεRI相比的损失不成比例的损失为 出乎意料，因为IE7不会导致MC在体外失去IgE，并且该MV AB仅应交联FcεRI，如果 通过聚集或与细胞Fcγ或补体受体结合，AB成为多价。 该提案以这些观察为基础。 AIM 1使用流式细胞术，结构研究，抗FCγRmAb， MV IE7的Hu IgG同种型开关变体，以确定MV HUIGG1 IE7是否在体内交叉链接FCεRI 如果是这样，则通过什么机制。 AIM 1还评估MV Huigg1 IE7聚合还是 IgE负载的FcεRI的内在化是MC IgE的体内损失不成比例所必需的。 AIM 2适用 AIM 1使用MV IE7安全地脱敏，表达表达的高敏感性高过敏性HUFCεRIα小鼠 突变体IL-4受体以及被动敏感的HU脐带血液恢复，hu细胞因子分泌的 免疫缺陷的小鼠会产生大量活化的HU MC和BAS；两种模型都是 由于二价ie7的剂量增加，很难脱敏。 AIM 3应用AIM 2结果以使恒河脱敏 猴子，表达与IE7反应的FcεRIα。这些目标在一起，应优化使用MV抗的使用 FcεRIαmAb用于RD，并建议是否以及如何使用它来抑制Hu IgE介导的疾病。

项目成果

Mucosal Nanoemulsion Allergy Vaccine Suppresses Alarmin Expression and Induces Bystander Suppression of Reactivity to Multiple Food Allergens.

• DOI: 10.3389/fimmu.2021.599296
• 发表时间: 2021
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Farazuddin M;Landers JJ;Janczak KW;Lindsey HK;Finkelman FD;Baker JR Jr;O'Konek JJ
• 通讯作者: O'Konek JJ