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

项目总结 包括结膜在内的眼部粘膜免疫动态平衡的破坏会导致疾病 以及潜在的视力受损。结膜是眼表面的一种富含免疫功能的粘膜组织。 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病毒药物来预防疾病。 微生物。结膜内的炎症细胞和细胞因子可以调节结膜的生长和随后的生长。 与感染性眼病相关的发病机制。已有研究表明，眼部微生物群可能 增强这一过程，从而在眼科疾病中发挥突出作用。清楚地定义了眼部微生物区系 中介性眼球免疫对于开发更有效的眼病治疗方法至关重要。我的 初步数据表明，共生体在眼粘膜内扩大了γδT细胞的一个亚群，这有助于 炎性单核细胞向眼粘膜的迁移及眼内抗微生物分子的释放 泪水。拟议研究的总体目标是确定眼部共生是如何诱导γδT细胞的 并确定这种特定的相互作用是否真的限制了传染性眼病。总体假设是 眼部γδT细胞是由共生体以抗原特异性的方式产生和维持的，这反过来又 保护眼表免受致病性耐甲氧西林金黄色葡萄球菌(MRSA)的感染 感染。计划中的研究的基本原理是，在完成后，眼睛和眼睛之间的新见解 微生物群和粘膜免疫系统将被揭示，然后可以被利用来缓解 疾病。该计划的K99阶段和特定目标1将使用细胞培养和荧光显微镜来 确定眼部微生物区系如何刺激眼部γδT细胞。特异靶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 维持共生定向的γδT细胞。最后，在独立的R00阶段，具体而言 目的3，我将使用眼部耐甲氧西林金黄色葡萄球菌感染的体内模型来显示所描述的共生/γδT细胞 AXIS保护眼睛环境免受病原性感染。该计划的培训阶段将是 主要在雷切尔·卡斯皮博士(导师)的实验室进行。培训目标包括接收 罗伯特·法里斯博士(共同导师)和NEI生物成像的荧光显微镜高级培训 CORE用于成像T细胞和抗原提呈细胞(APC)在原位和细胞培养中的相互作用。我 还将接受桑迪普·达塔博士(共同导师)和他的实验室的培训，以了解 与细菌感染有关的发病机制。来自拟议研究的数据将有助于定义 共生微生物区系调节眼睛免疫，这可能有助于眼病的治疗，同时也 为我终身从事眼科领域的独立研究奠定了基础 免疫学。

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.