Microbiome-dependent modulation of ocular immunity and the implications for infectious

微生物组依赖性眼部免疫调节及其对感染的影响

• 批准号: 9883003
• 负责人: ANTHONY J ST LEGER
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-04 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9883003
• 关键词: Affect; Antigen-Presenting Cells; Antigens; Area; Award; Bacteria; Bacterial Infections; Biological; Blindness; Cell Communication; Cell Culture Techniques; Cells; Conjunctivitis; Corynebacterium; DNA; Data; Development; Disease; Environment; Evidence based intervention; Experimental Designs; Eye diseases; Fluorescence Microscopy; Foundations; Generations; Goals; Graft Rejection; Homeostasis; Hour; Human; Hypersensitivity; Image; Immune; Immune response; Immune system; Immunity; Immunology; In Situ; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-17; Intervention; Keratitis; Keratoplasty; Laboratories; Life; Literature; Lymphocyte; Maintenance; Mediating; Mentors; Mucosal Immune System; Mucosal Immunity; Mucous Membrane; Mus; Organism; Pathogenesis; Pathogenicity; Patients; Pattern recognition receptor; Phase; Play; Positioning Attribute; Predisposition; Process; Recurrence; Regulation; Reporting; Research; Research Personnel; Role; Sjogren' s Syndrome; Staphylococcus aureus infection; T cell response; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Time; Tissues; Training; Visual impairment; antimicrobial; career; clinical development; commensal bacteria; commensal microbes; conjunctiva; cytokine; design; effective therapy; eye dryness; in vivo; in vivo Model; insight; methicillin resistant Staphylococcus aureus; microbiome; microbiota; microorganism; migration; monocyte; new therapeutic target; novel; ocular microbiome; ocular surface; pathogen; pathogenic bacteria; pathogenic microbe; prevent; programs; response; γδ T cells

PROJECT SUMMARY Disruption of ocular immune homeostasis in ocular mucosa, which includes the conjunctiva, leads to disease and potential impaired vision. The conjunctiva is a mucosal tissue of the ocular surface that is rich in immune cells. These immune cells can prevent disease by detecting antigen, producing cytokines, and secreting anti- microbials. Inflammatory cells and cytokines within the conjunctiva can regulate the outgrowth and subsequent pathogenesis associated with infectious ocular disease. It has been suggested that the ocular microbiome may augment this process, thus playing a prominent role in ocular disease. Clearly defining how ocular microbiota mediate ocular immunity will be crucial for the development of more effective therapies for ocular diseases. My preliminary data suggest that commensals expand a subset of γδ T cells within ocular mucosa, which facilitate the migration of inflammatory monocytes into ocular mucosa and the release of anti-microbial molecules within the tears. The overall goals of the proposed studies are to define how ocular commensals induce γδ T cells and to determine if this specific interaction truly limits infectious ocular disease. The overall hypothesis is that ocular γδ T cells are generated and maintained by commensals in an antigen-specific manner, which in turn protects the ocular surface from infection by pathogenic methicillin-resistant Staphylococcus aureus (MRSA) infection. Rationale for the research planned is such that at its completion, new insights between the ocular microbiome and mucosal immune system will be revealed, which can then be exploited for the alleviation of disease. The K99 PHASE of this plan and Specific Aim 1 will use cell culture and fluorescence microscopy to identify how ocular γδ T cells are stimulated by ocular microbiota. Specific Aim 2 will use in vitro and in vivo techniques to assess whether innate pattern recognition receptors (PRRs) contribute to the generation and/or maintenance of commensal-directed γδ T cells. Lastly, during the INDEPENDENT R00 PHASE, in Specific Aim 3, I will use an in vivo model of ocular MRSA infection to show whether the described commensal/γδ T cell axis protects the ocular environment from pathogenic infection. The training phase of this plan will be conducted mainly in the laboratory of Dr. Rachel Caspi (mentor). TRAINING OBJECTIVES include receiving advanced training in fluorescence microscopy by Dr. Robert Fariss (co-mentor) and the NEI biologic imaging core for the purpose of imaging T cell and antigen presenting cell (APC) interactions in situ and in cell culture. I will also receive training from Dr. Sandip Datta (co-mentor) and his laboratory for the purpose of understanding pathogenesis associated with bacterial infections. Data from the proposed studies will help define how commensal microbiota modulate ocular immunity, which may aid in the treatment of ocular disease while also providing me a foundation for a life-long career as an independent researcher in the field of ocular immunology.

项目摘要 包括结膜在内的眼粘膜中的眼免疫稳态的破坏导致疾病 以及潜在的视力受损结膜是眼表的粘膜组织，其富含免疫球蛋白。 细胞这些免疫细胞可以通过检测抗原、产生细胞因子和分泌抗 微生物结膜内的炎性细胞和细胞因子可以调节结膜的生长和随后的炎症反应。 与感染性眼病相关的发病机制。有人认为眼部微生物组可能 增强这一过程，从而在眼部疾病中发挥突出作用。清楚地定义了眼部微生物群 介导的眼部免疫对于开发更有效的眼部疾病治疗方法至关重要。我 初步数据表明，眼内分泌系统扩增了眼粘膜内的γδ T细胞亚群， 炎性单核细胞向眼粘膜的迁移和眼粘膜内抗微生物分子的释放 眼泪这些研究的总体目标是确定眼内分泌物如何诱导γδ T细胞 并确定这种特定的相互作用是否真正限制了传染性眼病。总的假设是， 眼γδ T细胞是由免疫球蛋白以抗原特异性的方式产生和维持的， 保护眼表免受致病性耐甲氧西林金黄色葡萄球菌（MRSA）感染 感染计划进行研究的理由是，在完成研究时， 微生物组和粘膜免疫系统将被揭示，然后可以用于缓解 疾病本计划的K99阶段和具体目标1将使用细胞培养和荧光显微镜， 确定眼部γδ T细胞如何被眼部微生物群刺激。Specific Aim 2将在体外和体内使用 评估先天模式识别受体（PRR）是否有助于产生和/或 维持细胞介导的γδ T细胞。最后，在独立R 00阶段，具体而言， 目的3，我将使用眼部MRSA感染的体内模型来显示所描述的结膜/γδ T细胞是否被抑制。 轴保护眼环境免受病原体感染。该计划的培训阶段将是 主要在Rachel Caspi博士（导师）的实验室进行。培训内容包括： Robert Fariss博士（共同导师）和NEI生物成像的荧光显微镜高级培训 核心，用于原位和细胞培养中对T细胞和抗原呈递细胞（APC）相互作用进行成像。我 还将接受Sandip Datta博士（共同导师）及其实验室的培训，以了解 与细菌感染相关的发病机制。来自拟议研究的数据将有助于确定 眼部微生物群调节眼部免疫，这可有助于治疗眼部疾病，同时还 为我作为一个独立的研究人员在眼科领域的终身职业生涯奠定了基础， 免疫学