Plasma Serum Based Biomarkers in Sublingual Oral Immunotherapy for Milk Allergy

基于血浆血清的生物标志物用于舌下口服免疫疗法治疗牛奶过敏

• 批准号: 8424318
• 负责人: JOHN T. SCHROEDER
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-15 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424318
• 关键词: Accounting; Acids; Address; Adolescence; Adult; Adverse reactions; Affect; Aftercare; Allergen Immunotherapy; Allergens; Allergic; Antibodies; Area; Basophils; Biological Assay; Biological Markers; Blood; CD34 gene; Cattle; Child; Clinical; Complex; Data; Dendritic Cells; Development; Diet; Disease; Effectiveness; Elements; Food; Food Contamination; Food Hypersensitivity; Food Processing; Future; Goals; Hand; Histamine; Histamine Release; Hypersensitivity; IgE; IgG4; Immune response; Immunoglobulin G; Immunoglobulins; Immunotherapy; In Vitro; Individual; Institution; Interferons; Interleukin-4; Link; Liquid substance; Mediating; Milk; Milk Hypersensitivity; Milk Proteins; Monitor; Oral; Patient Care; Patients; Pilot Projects; Plasma; Play; Pre-Post Tests; Process; Production; Reaction; Recording of previous events; Relative (related person); Reporting; Research Project Grants; Restaurants; Risk; Role; Safety; Sampling; Serum; Specimen; Testing; Time; Work; adult stem cell; airborne allergen; assay development; base; clinical efficacy; clinically relevant; crosslink; cytokine; desensitization; effective therapy; follow-up; food allergen; human SYK protein; in vitro testing; novel strategies; omalizumab; oral immunotherapy; passive sensitization; receptor; research study; response

DESCRIPTION (provided by applicant): Cow's milk allergy (CMA) is the most common food allergy in young children and is now likely to persist into adolescence and adulthood. Strict milk avoidance remains the only treatment for this allergy but accidental reactions, which are often severe, are extremely common, frequently due to the hidden presence of food allergens in processed foods and cross-contamination of foods. Recent studies from our institution and elsewhere have identified oral and sublingual allergen immunotherapy (OIT and SLIT) as promising approaches for treatment of CMA. Studies demonstrate that most children with IgE-mediated CMA can be desensitized to cow's milk protein, both safely and efficaciously. However, the specific mechanisms by which this treatment appears to work remain unknown, and questions remain to whether this increased tolerance to milk after treatment is permanent, or rather represents a transient desensitization which could place patients at significant risk of future reactions if milk was ever discontinued from their diet. Our long-range goal is to develop in vitro tests that help predict risks to adverse reactions to food as well as the safety/clinical efficacy of OIT/SLIT. The best chance of developing a routine test to make such predictions is best achieved with the versatility that serum/plasma provides. Our novel approach of using culture-derived basophils (CDBA) grown from adult stem cells should facilitate identification of plasma markers. Therefore, the proposal consists of 2 aims: Aim 1 involves analyzing plasma from CMA children undergoing SLIT and OIT for "blocking activity" by comparing pre and post specimens in passively sensitizing CDBA for subsequent challenge with milk allergen. The advantage of using CDBA is that they have never been exposed to immunoglobulins (IgE/IgG), which may otherwise confound the ability to detect blocking IgG4 antibody. Similar experiments will test pre- and post- OIT/SLIT plasma specimens for effects on plasmacytoid dendritic cells (pDCs), predicting that post-OIT/SLIT plasma (with greater IgG4 levels) will augment innate immune responses (TLR9) that are seemingly impaired in allergic individuals. Aim 2 explores the existing hypothesis that circulating allergen-IgE complexes are in plasma of CMA subjects and that these can be detected by inducing phenotypic/functional responses in CDBA and/or normal basophil. Plasma specimens from CMA subjects will be tested in flow-based assays for their ability to induce basophil spontaneous histamine release (SHR), the expression of markers (CD63/CD203c) linked to IgE-dependent activation as well as those (syk kinase) that are down regulated with prolonged Fc5RI cross-linking. These assays could result in new plasma-based tests useful in predicting whether one is at risk for adverse reactions and for predicting the clinical efficacy during successful OIT/SLIT. With food specific immunotherapy being potentially the most exciting area in the history of food allergy research, this project, with clinical and mechanistic endpoints should have immediate impact on the care of patients with food allergy.

描述（由申请人提供）：牛奶过敏（CMA）是幼儿中最常见的食物过敏，现在很可能会持续到青春期和成年期。严格的避免牛奶仍然是这种过敏的唯一治疗方法，但是意外反应通常很严重，这是极为普遍的，这通常是由于食物过敏原的隐藏存在和食物的交叉污染。我们机构和其他地方的最新研究确定了口服和舌下过敏原免疫疗法（OIT和SLIT）是CMA治疗的有希望的方法。研究表明，大多数患有IgE介导的CMA的儿童可以安全和有效地将其脱敏牛奶蛋白。但是，这种治疗方法似乎起作用的具体机制仍然未知，并且对治疗后对牛奶的耐受性的提高仍然存在问题，或者代表了一种短暂的脱敏，如果牛奶曾被饮食中断，可能会使患者处于未来反应的危险中。我们的远程目标是开发体外测试，以帮助预测对食物的不利反应以及OIT/SLIT的安全/临床功效。通过血清/等离子体提供的多功能性，可以最好地制定常规测试来做出这种预测的最佳机会。我们使用成人干细胞种植的培养衍生的嗜碱性粒细胞（CDBA）的新方法应促进血浆标记的鉴定。因此，该提案由2个目标组成：AIM 1涉及分析接受SLIT和OIT的CMA儿童的血浆，以通过比较样品前后的样品被动地将CDBA进行比较，以使CDBA与牛奶过敏原一起挑战。使用CDBA的优点是它们从未暴露于免疫球蛋白（IgE/IgG），否则它们可能会混淆检测阻断IgG4抗体的能力。类似的实验将测试OIT/SLIT血浆标本对浆细胞类动物树突状细胞（PDC）的影响，预测OIIT/SLIT血浆（具有较高的IgG4水平）将增加天生的免疫反应（TLR9），这似乎受到了镇压个体的影响。 AIM 2探讨了现有的假设，即CMA受试者的血浆中循环过敏蛋白 - IGE复合物，可以通过在CDBA和/或正常嗜碱性粒细胞中诱导表型/功能反应来检测这些假设。来自CMA受试者的血浆标本将在基于流量的测定中进行测试，以诱导basspophil自发性组胺释放（SHR），标记物（CD63/CD203C）与IgE依赖性激活相关的标记（CD63/CD203C）的表达以及（SYK激酶）（SYK激酶）被延长的FC5RI交叉链接下调。这些测定可能导致新的基于等离子体的测试有助于预测一个人是否有不良反应的风险以及预测成功的OIT/SLIT期间的临床疗效。由于食物特定的免疫疗法可能是食品过敏研究史上最令人兴奋的领域，因此具有临床和机械终点的项目应立即对食物过敏患者的护理产生直接影响。