The Role of Innate Immune Cell Trafficking in Th2 Differentiation

项目摘要 候选 卡罗琳索科尔，医学博士，博士，是过敏和临床免疫学部门的医学助理， 马萨诸塞州总医院（MGH）和哈佛医学院（HMS）医学讲师。她 获得医学博士学位/博士来自耶鲁大学，在那里她研究了先天免疫控制的机制， Ruslan Medzhitov博士实验室中的Th 2分化。她在那里的工作导致了两个第一作者自然 关于嗜碱性粒细胞在引发过敏性免疫应答和Th 2中的作用的免疫学出版物 分化这也导致了她目前在中心的安德鲁·卢斯特博士实验室的工作。 免疫学和炎症性疾病（CIID）在MGH，在那里她专注于先天免疫的作用， 在过敏性免疫应答和Th 2分化的激活和控制中的细胞运输。短- 这个K 08奖申请重新提交的长期目标是提供活体显微镜的培训， 先进的基因组技术，包括RNA-Seq，人类免疫学和转化研究。索科尔医生 长期目标是发展一个独立的转化研究计划，研究先天免疫控制 过敏性疾病本提案中概述的实验、培训活动和指导计划将 成功地定位索科尔博士为她第一个R 01和一个独立的职业生涯作为一个物理学家，科学家。 指导、培训活动和环境 博士索科尔将在MGH的CIID中执行本提案中概述的工作，并接受Dr. 安德鲁·卢斯特CIID是一个最先进的多学科基础科学研究中心，专注于 作为免疫学基础的免疫介导的炎症性疾病的机制 研究在MGH。Luster博士是细胞运输和趋化因子研究领域的专家， 是这个应用程序的核心。此外，Luster博士有着出色的指导记录， 指导了50多名博士后候选人，其中17人目前是独立资助的研究学院， 学术机构此外，索科尔博士还组织了一个培训咨询委员会， 尼尔·哈科恩，托尔斯滕·曼佩尔和阿琳·夏普。他们将提供专业知识和实践培训， 先进的显微成像，先进的基因组技术，人类免疫学和转化研究。 为了补充导师的专业知识，索科尔博士将完成生物信息学的教学课程， 系统生物学，资助写作，领导力，转化研究和负责任的研究行为 通过哈佛催化剂。合作机会、知识环境和资源 索科尔博士所能获得的都是杰出的，并为她提供了一条通往独立成功的明确道路。 研究 过敏性疾病的特征在于针对非感染性免疫应答的不适当的2型免疫应答。 环境过敏原尽管过敏性疾病的发病率很高，但人们对过敏性疾病是如何发生的知之甚少。 先天免疫系统识别过敏原并启动过敏性免疫应答， 研究新型疗法。最近的研究已经确定了一个专门的CD 301 b+树突状细胞群， 位于鼠皮肤中的DC细胞（DCs），其选择性地迁移到引流淋巴结（dLN）中并促进 Th 2细胞对过敏原的反应。然而，促进这种选择性迁移的途径， 对过敏原的反应尚不清楚。我们建议，首先，定义参与的趋化因子途径， CD 301 b + DC向dLN的迁移。我们在初步数据中显示，过敏原暴露导致 CCR 8配体在小鼠皮肤和dLN以及人皮肤外植体中的产生。而且我们 提供了初步数据，除了CCR 7信号外，CCR 8对于CD 301 b+的迁移是必需的。 CCR 2阻止CD 301 b + DC从皮肤迁移到皮肤，随后分化为Th 2，而CCR 2阻止CD 301 b + DC迁移到皮肤。 dLN。然而，CD 301 b + DC迁移的控制并不是先天性控制过敏性疾病的唯一机制。 免疫启动尽管CD 301 b + DC对于Th 2分化是必需的，但它们不足以 诱导幼稚T细胞Th 2分化。接下来，我们建议确定 辅助细胞促进Th 2分化。我们提供的初步数据表明， 辅助细胞，提供与抗原呈递串联的必要偏斜信息， DC的共刺激。这些辅助细胞通过CD 62 L依赖性过程进入dLN， 该细胞类型与我们已经鉴定的先前未描述的产生IL-4的细胞相对应。 我们认为，通过控制CD 301 b + DC的迁移和这种辅助细胞的进入， 先天免疫系统精确地控制T辅助细胞分化的结果。

PROJECT SUMMARY Candidate Caroline Sokol, M.D., Ph.D., is an Assistant in Medicine in the Allergy and Clinical Immunology Unit at Massachusetts General Hospital (MGH) and an Instructor of Medicine at Harvard Medical School (HMS). She received her M.D./Ph.D. from Yale University, where she studied the mechanisms of innate immune control of Th2 differentiation in the laboratory of Dr. Ruslan Medzhitov. Her work there led to two first author Nature Immunology publications on the role of basophils in initiating the allergic immune response and Th2 differentiation. It also led to her current work in the laboratory of Dr. Andrew Luster at the Center for Immunology and Inflammatory Diseases (CIID) at MGH, where she has focused on the role of innate immune cell trafficking in the activation and control of the allergic immune response and Th2 differentiation. The short- term goals of this K08 award application resubmission are to provide training in intravital microscopy, advanced genomic techniques, including RNA-Seq, human immunology and translational research. Dr. Sokol’s long-term goal is to develop an independent translational research program studying the innate immune control of allergic disease. The experiments, training activities, and mentoring plan outlined in this proposal will successfully position Dr. Sokol for her first R01 and an independent career as a physician-scientist. Mentorship, Training Activities, and Environment Dr. Sokol will perform the work outlined in this proposal in the CIID at MGH, under the mentorship of Dr. Andrew Luster. The CIID is a state-of-the-art multidisciplinary basic science research center focused on mechanisms of immune-mediated inflammatory diseases that serves as the foundation for immunology research at MGH. Dr. Luster is an expert in the fields of cellular trafficking and chemokine research, areas that are central to this application. Additionally, Dr. Luster has an excellent track record of mentorship, having mentored over 50 post-doctoral candidates, of which 17 are currently independently-funded research faculty at academic institutions. Additionally, Dr. Sokol has organized a Training Advisory Committee consisting of Drs. Nir Hacohen, Thorsten Mempel and Arlene Sharpe. They will provide expertise and hands-on training in advanced microscopic imaging, advanced genomic techniques, human immunology and translational research. To complement the expertise of her mentors, Dr. Sokol will complete didactic courses in bioinformatics and systems biology, grant writing, leadership, translational research, and the responsible conduct of research through the Harvard Catalyst. The collaborative opportunities, intellectual environment, and resources available to Dr. Sokol are outstanding and provide her with a clear path to independent success. Research Allergic diseases are characterized by inappropriate Type-2 immune responses targeted against non-infectious environmental allergens. Despite the high prevalence of allergic diseases, very little is known about how the innate immune system recognizes allergens and initiates the allergic immune response, which has limited research into novel therapeutics. Recent research has identified a specialized population of CD301b+ dendritic cells (DCs) located in murine skin that selectively migrates into the draining lymph node (dLN) and promotes Th2 differentiation in response to allergens. However, the pathways that promote this selective migration in response to allergens are unknown. We propose first, to define the chemokine pathways involved in the migration of CD301b+ DCs to the dLN. We show in preliminary data that allergen exposure results in the production of CCR8 ligands in the skin and dLN of mice, as well as in human skin explants. Furthermore, we provide preliminary data that in addition to CCR7 signals, CCR8 is necessary for the emigration of CD301b+ DCs from the skin and subsequent Th2 differentiation, while CCR2 prevents CD301b+ DC emigration to the dLN. However, control of CD301b+ DC migration is not the only mechanism for innate control of allergic immune initiation. Although CD301b+ DCs are necessary for Th2 differentiation, they are not sufficient to induce Th2 differentiation from naïve T cells. We next propose to determine the role and identity of accessory cells in promoting Th2 differentiation. We provide preliminary data that there is a required accessory cell that provides the necessary skewing information in tandem with antigen presentation and costimulation by DCs. These accessory cells enter the dLN via a CD62L-dependent process and the entry of this cell type corresponds with the entry of a previously undescribed IL-4 producing cell that we have identified. We believe that through the control of CD301b+ DC migration and the entry of this accessory cell, that the innate immune system precisely controls the outcome of T helper cell differentiation.