Understanding the microbial requirements for colonization and immunogenicity of commensal bacteria at the ocular surface

摘要 眼表疾病每年造成超过120亿美元的经济负担，令人震惊 治疗。而目前的护理标准只治标不治本(S) 对于疾病，最近的数据表明，操纵微生物组可能是一种潜在的途径 治疗和/或预防疾病。最近，一种眼共生细菌，棒状杆菌 Mastidis(C.mast)被发现并被证明能保护眼表免受C. 白色念珠菌和铜绿假单胞菌。尽管有这种有益的豁免权，但仍有必要了解 控制眼球定植和刺激宿主免疫能力的微生物因素 回应。拟议研究的基础是从以下方面获得的初步数据 30多株棒状杆菌临床相关菌株的基因组和表型筛选 SPP.和近2000个转座子突变体。具体地说，第一个目标将确定 控制眼睛定植的微生物因素(S)，通过测试8个突变的桅杆弯曲菌 预计会缺乏定植眼睛的能力。第二个目标将发现微生物因素(S) 通过评估九种不同的C. 肥大突变体被预测具有有限的诱导免疫反应的能力。第三 AIM将从机械上定义眼部共生细菌如何刺激人类免疫 回应。正如所概述的，拟议的研究将提供多个层面的有益信息 为了眼睛健康。首先，目标1和目标2将提供发挥关键作用的特定基因的身份 在眼睛共生的性质中的作用，这将使未来的研究更好地区分 暂时进入眼睛的其他细菌的眼部共生体。这些目标 也将阐明在转基因眼睛的配方中可能需要的基因 细菌或眼部益生菌。最后，目标3将提供关于一个几乎未知的有价值的知识 区域，这是人类对棒状杆菌的免疫反应。因为 棒状细菌在人体内相当普遍，这些数据将产生深远的影响 不仅对眼睛，而且对身体的其他部位都有影响。总括而言，拟议的研究 将为未来眼部益生菌治疗眼部疾病的发展奠定基础。

Abstract Ocular surface disease has an astounding economic burden of over $12 billion per year for treatment. While current standards of care only treat symptoms rather than address the cause(s) of disease, recent data suggest that manipulation of the microbiome may be a potential avenue to treat and/or prevent disease. Recently, an ocular commensal bacterium, Corynebacterium mastitidis (C. mast), was identified and shown to protect the ocular surface from infection with C. albicans and P. aeruginosa. Despite this beneficial immunity, there is still a need to understand the microbial factors that govern ocular colonization and ability to stimulate the host immune response. The foundation of the proposed studies was built upon preliminary data acquired from genomically and phenotypically screening over 30 clinically relevant isolates of Corynebacterium spp. and nearly 2000 transposon mutants of C. mast. Specifically, the first aim will identify microbial factor(s) that govern ocular colonization by testing eight C. mast mutants that were predicted to lack an ability to colonize the eye. The second aim will discover microbial factor(s) that stimulate host immunity by assessing immunogenicity, in vitro and in vivo, of nine different C. mast mutants that were predicted to have a limited ability to induce immune responses. The third aim will mechanistically define how ocular commensal bacteria stimulate the human immune response. As outlined, the proposed studies will provide multiple layers of information beneficial to ocular health. First, Aims 1 and 2 will provide the identity of specific genes that play a critical role in the nature of an ocular commensal, which will allow future studies to better distinguish true ocular commensals from other bacteria that are temporarily introduced to the eye. These aims will also shed light on genes that may be desirable in the formulation of genetically modified ocular bacteria or ocular probiotics. Finally, Aim 3 will provide valuable knowledge on a virtually unknown area, which is the human immune response against Corynebacterium spp. Because Corynebacteria are fairly ubiquitous throughout the body, these data will have far-reaching implications for not only the eye, but throughout the rest of the body. In sum, the proposed studies will form the foundation for the future development of ocular probiotics to treat eye diseases.