Humoral mechanisms of tolerance in peanut oral immunotherapy

 应聘者：Sarita Patil，医学博士，马萨诸塞州总医院过敏和免疫学专科医生，哈佛医学院医学讲师，初级研究员。在她担任研究员期间，她进行了一项转化研究，这导致了一种荧光过敏原特异性B细胞四聚体的开发，该四聚体可以成功地识别正在接受免疫治疗的花生过敏患者周围的花生特异性B细胞。在这项技术的基础上，这款K23应用程序旨在了解体液反应在口腔食物免疫治疗中的作用，特别是在克隆水平上研究免疫球蛋白对花生过敏原Ara h2的反应，从而更深入地了解人类食物过敏患者的免疫治疗机制。拟议的项目结合了临床试验、基础科学和生物信息学的研究，以开发翻译研究方面的独特专业知识。本提案中概述的工作、培训活动和指导计划将成功地将Patil博士定位为HR First R01应用程序，并成为一名独立的内科科学家。指导、培训活动和环境：Patil博士将在MGH免疫学和炎症性疾病中心(CIID)和麻省理工学院(MIT)分别在Wayne Shreffler博士和J.Christopher Love博士的共同指导下执行拟议的项目。CIID是一个多学科的基础科学中心，是麻省理工学院免疫学研究的基础。此外，麻省理工学院的罗伯特·安东尼博士和耶鲁大学医学院的史蒂文·克莱因斯坦博士将在帕蒂尔博士的咨询委员会任职，分别提供过敏性疾病中免疫球蛋白糖基化以及使用计算生物学方法进行免疫球蛋白图谱分析方面的专业知识。为了补充她提出的项目和导师的专业知识，帕蒂尔博士将通过哈佛公共卫生学院和哈佛大学催化剂完成翻译研究、统计学、测序、蛋白质组学和生物信息学方面的授课课程。帕蒂尔博士所拥有的合作机会、智力环境和资源都非常出色。研究：免疫球蛋白介导的花生过敏对公共健康构成重大风险，因为接触微小的花生会导致严重的过敏反应。在仔细观察下，花生口服免疫疗法(PNOIT)在儿童中逐渐增加花生暴露的临床试验已经成功地诱导了脱敏；然而，这些儿童中只有一小部分具有长期耐受性。接受PNOIT治疗的患者通常花生特异性IgE降低，花生特异性免疫球蛋白和IgG4水平逐渐升高，但这些花生特异性免疫球蛋白变化对长期耐受的机制贡献仍不清楚。这个项目解决了PNOIT如何影响B细胞谱系，以及这些变化如何与预期的不同临床结果相关。我们假设，PNOIT后的长期临床耐受性与高亲和力和寡克隆性变应原特异性功能性抗体的诱导直接相关，并部分归因于此。我们希望使用基于荧光四聚体的方法来鉴定变应原特异性B细胞，以确定临床预后标志物，并提高我们对免疫治疗机制的理解，特别是抗体的病理生理作用及其作为耐受生物标志物和治疗药物的潜力。为此，我们建议在PNOIT患者中使用蛋白质组学和下一代测序来定位过敏原特异性B细胞谱系。我们还建议对来自过敏原特异性B细胞的重组抗体进行功能研究，以探索PNOIT耐受的潜在机制。通过对PNOIT期间产生可变耐受性的花生过敏患者和自然不再对其过敏的花生过敏患者的研究，我们希望阐明B细胞系在花生过敏中诱导长期耐受中的作用。这些努力可能有助于确定相关的临床生物标志物和潜在的治疗途径，以改进PNOIT治疗花生过敏的效果。

 DESCRIPTION (provided by applicant): Candidate: Sarita Patil, MD is a staff physician in Allergy and Immunology at Massachusetts General Hospital (MGH) and a junior investigator as an Instructor in Medicine at Harvard Medical School (HMS). During her fellowship, she conducted translational research, which led to the development of a fluorescent allergen- specific B cell tetramer that could successfully identify peanut-specific B cells in the periphery f peanut- allergic patients undergoing immunotherapy. Building on that technology, this K23 application aims to understand the role of the humoral response in oral food immunotherapy, specifically investigating the immunoglobulin response to the peanut allergen Ara h 2 on a clonal level, allowing for greater insight into mechanisms of immunotherapy in human patients with food allergy. The proposed project combines research in clinical trials, basic science, and bioinformatics to develop a unique expertise in translational research. The work, training activities, and mentoring plan outlined in this proposal will successfully position Dr. Patil for hr first R01 application and an independent career as a physician-scientist. Mentorship, Training Activities, and Environment: Dr. Patil will perform the proposed project in the Center for Immunology and Inflammatory Diseases (CIID) at MGH under the mentorship of Dr. Wayne Shreffler and at Massachusetts Institute of Technology (MIT) under the co-mentorship of Dr. J. Christopher Love. The CIID is a multidisciplinary basic science center that serves as the foundation of immunology research at MGH. In addition, Dr. Robert Anthony at MGH and Dr. Steven Kleinstein at Yale School of Medicine will serve on Dr. Patil's Advisory Committee to provide expertise on immunoglobulin glycosylation in allergic diseases as well as immunoglobulin repertoire analysis using computational biology approaches, respectively. To complement her proposed projects and the expertise of her mentors, Dr. Patil will complete didactic courses in translational research, statistics, sequencing, proteomics, and bioinformatics through the Harvard School of Public Health and the Harvard Catalyst. The collaborative opportunities, intellectual environment and resources available to Dr. Patil are outstanding. Research: IgE-mediated peanut allergy poses a significant public health risk, as minute peanut exposures can cause severe allergic reactions. Clinical trials of peanut oral immunotherapy (PNOIT) with gradual incremental peanut exposure under careful observation in children have been successful in inducing desensitization; however, only a fraction of these children have long-lived tolerance. Patients undergoing PNOIT often have a decrease in their peanut-specific IgE and a gradual increase in their peanut-specific IgG and IgG4 levels, but the mechanistic contribution of these changes in peanut-specific immunoglobulins to long-term tolerance is still unknown. This project addresses how PNOIT affects the B cell repertoire and how those changes correlate to the heterogeneous clinical outcomes that are expected. We hypothesize that long-lasting clinical tolerance following PNOIT directly correlates with, and is partially dueto the induction of high affinity and oligoclonal allergen-specific functional antibodies. Using a fluorescent tetramer-based approach for the identification of allergen-specific B cells, we hope to identify clinical prognostic markers, and improve our understanding of the mechanism of immunotherapy, specifically the pathophysiological role of antibodies and their potential as both biomarkers of tolerance and therapeutic agents. To do so, we propose mapping the allergen-specific B cell repertoire using proteomics and next- generation sequencing in PNOIT patients. We also propose functional studies of recombinant antibodies from the allergen-specific B cells to explore the underlying mechanism of tolerance in PNOIT. By studying both peanut allergic patients who develop variable tolerance during PNOIT as well as peanut allergic patients who naturally outgrew their allergies, we hope to elucidate the contribution of the B cell repertoire t the induction of long-lived tolerance in peanut allergy. These efforts may help identify relevant clinical biomarkers and potential therapeutic avenues for the improvement of PNOIT for the treatment of peanut allergy.