Pathogenesis of Uveitis in Ebola Virus Disease Survivors

项目摘要/摘要 埃博拉病毒病(EVD)与埃博拉病毒病幸存者中葡萄膜炎的高发病率有关 2014-2016年间西非史无前例的埃博拉疫情。疾病的范围从 前葡萄膜炎到威胁视力的全葡萄膜炎，完全失明。埃博拉病毒幸存者的绝对数量 IMPACTIVE给了我们独特的机会来描述临床特征，结构性并发症 导致视力丧失，以及葡萄膜炎的发病机制。除了视力丧失对视力质量的显著影响外 日常生活活动中，与埃博拉病毒(EBOV)相关的葡萄膜炎具有科学性和公共卫生性 可能是因为我们之前在房水中发现了EBOV。除了最近爆发的 西非，刚果民主共和国最近的两起疫情突显了全球卫生 授权充分了解埃博拉病毒病后遗症的临床和科学影响。 新兴的临床、基础和转化性研究提供了关于潜在的洞察力 葡萄膜炎与EVD的相关机制。我们最近描述了我们的新方法，以安全地 询问EVD幸存者的房水标本以了解EBOV；同时检测这些眼液标本 一例非人类灵长类动物眼组织的最新分子病理学分析 EVD幸存者模型已经证明EBOV持续存在于巨噬细胞的玻璃体内，并且是 与葡萄膜炎和视网膜炎有关。一名患有葡萄膜炎的埃博拉病毒病幸存者的更多引人注目的免疫学研究 显示埃博拉特异性T细胞和B细胞激活，血浆后有持续的埃博拉抗原刺激迹象 EBOV的通关。这些发现暗示了EBOV的长期持续，这是可以通过 全身性T和B细胞反应。为了进一步描述葡萄膜炎的患病率和临床谱系， 对于EVD幸存者的EBOV持久性和葡萄膜炎的免疫学机制，我们提出了三个目标： 1)在目标1中，将比较埃博拉病毒病幸存者和塞拉利昂密切接触者之间葡萄膜炎的患病率 里昂。将评估临床因素和宿主风险因素，包括人类白细胞抗原分型，以确定是否有 葡萄膜炎发生的临床和宿主相关决定因素。 2)在AIM 2中，EBOV的持久性将使用逆转录聚合酶链式反应(RT- 玻璃体液的聚合酶链式反应检测)和原位杂交技术检测玻璃体中的EBOV基因组。 3)在目标3中，将评估埃博拉幸存者的血液和眼液中的埃博拉特异性免疫反应 对于葡萄膜炎，专门评估埃博拉特异性B细胞和T细胞、免疫球蛋白亚类组成的激活情况。 这项研究的见解将定义与EVD相关的葡萄膜炎的频谱，评估EBOV是否 ，并促进我们对感染和免疫之间相互作用的理解 埃维德。最终，这些发现将确定EBOV的诊断策略和未来的医学对策， 以及可以在独特的免疫特权眼睛环境中建立持久性的其他病毒。

Project Summary/ Abstract Ebola virus disease (EVD) has been associated with a high prevalence of uveitis in EVD survivors in the wake of the unprecedented West African EVD outbreak from 2014-2016. The spectrum of disease ranges from anterior uveitis to sight-threatening panuveitis with complete blindness. The sheer number of EVD survivors impacted has given us the unique opportunity to characterize the clinical features, structural complications leading to vision loss, and pathogenesis of uveitis. Besides the significant impact of vision loss on quality-of- life and activities of daily living, uveitis associated with Ebola virus (EBOV) has scientific and public health implications because of our prior identification of EBOV in the aqueous humor. Beyond the recent outbreak in West Africa, two more recent outbreaks in Democratic Republic of Congo underscore the global health mandate to fully understand the clinical and scientific implications of EVD sequelae. Emerging clinical, basic, and translational investigation has provided insight regarding the potential mechanisms of uveitis associated with EVD. We recently described our novel methodology to safely interrogate aqueous humor specimens for EBOV in EVD survivors; while these ocular fluid specimens tested negative for EBOV by RT-PCR, recent molecular pathologic analysis of ocular tissue in a non-human primate EVD survivor model has demonstrated that EBOV persists within the vitreous humor in macrophages, and is associated with uveitis and retinitis. Additional compelling immunologic studies of an EVD survivor with uveitis showed Ebola-specific T-cell and B-cell activation with signs of ongoing Ebola antigen stimulation after plasma clearance of EBOV. These findings were suggestive of long-term EBOV persistence, which are detectable by systemic T- and B-cell responses. To further characterize the prevalence and clinical spectrum of uveitis, role of EBOV persistence and immunologic mechanisms of uveitis in EVD survivors, we propose three Aims: 1) In Aim 1, the prevalence of uveitis will be compared between EVD survivors and close contacts in Sierra Leone. Clinical factors and host risk factors including HLA-typing will be assessed to determine if there are clinical and host-related determinants of uveitis development. 2) In Aim 2, EBOV persistence will be assessed using reverse transcriptase polymerase chain reaction (RT- PCR testing) of vitreous fluid and in situ hybridization techniques to detect EBOV genome in the vitreous. 3) In Aim 3, Ebola-specific immune responses will be assessed in the blood and ocular fluid of EVD survivors with uveitis, specifically evaluating activation of Ebola-specific B and T-cells, IgG subclass composition. Insights from this study will define the spectrum of uveitis associated with EVD, assess whether EBOV is harbored in the vitreous, and advance our understanding of the interplay between infection and immunity in EVD. Ultimately, these findings will define diagnostic strategies and future medical countermeasures for EBOV, as well as other viruses that may establish persistence in the unique immune privileged ocular environment.