Genomic systems approach to measure HSV-1-specific T-cell dominance in humans

描述（由申请方提供）：1型单纯疱疹病毒（HSV-1）是一种具有医学意义的病原体。60%的美国公民被感染，发展中国家的感染率更高。严重的并发症，如成人或新生儿脑炎造成的永久性脑损伤，以及导致失明和即使在角膜移植后长期预后不良的角膜炎症，发生率很低。由于感染是如此普遍，这些并发症的累积医疗影响是相当可观的。目前还没有获得许可的HSV-1疫苗。HSV-2是一种具有50%氨基酸同一性的病原体，其疫苗在最近的III期试验中没有临床活性。这种HSV-2疫苗靶向抗体和CD 4 T细胞应答，但不靶向CD 8 T细胞应答。本研究的长期目标是合理设计HSV-1的预防性和治疗性疫苗。CD 8 T细胞是对HSV-1的免疫应答的关键组分，并且定位于感染的皮肤、角膜、视网膜和三叉神经节。从人类HSV-2和鼠HSV-1数据推断，CD 8细胞在愈合的皮肤感染部位持续存在，以提供对再活化的早期监测。几乎没有人知道它们在人类身上的特异性。我们开发了新的技术，使我们能够以前所未有的细节研究CD 8和CD 4 T细胞对HSV-1的反应，包括研究从未有过疱疹病变的人的外周血，即免疫成功的人。目的1使用一系列新的方法：有效的交叉呈递，基于CD 137的分选，和克隆的HSV-1 ORF文库，以高通量和清晰度解码CD 8对HSV的反应。我们将比较有症状和无症状HSV-1感染者的免疫应答，并确定具有高人群患病率和与无症状感染相关的抗原。HSV-1特异性CD 4定位于人类感染的眼睛、TG和皮肤，以及小鼠感染的组织，并可能提供重要的直接抗病毒和辅助功能。目的2使用完整的ORFeome，以高通量定义HSV-1特异性CD 4应答的广度、特异性和受试者内真实免疫显性层次。该工作流程使用基于CD 137的分选和高度抗原性的HSV-1 ORFeome蛋白组。优势测定在两个维度上是组合的（应答细胞寡克隆性和基质ORF合并），并提供每个受试者约100个单个HSV-1特异性CD 4 T细胞的精细特异性，以定义CD 4优势结构。该项目的总体目标是鉴定HSV-1的ORF，用于刺激与轻度感染相关的不同HLA类型的人的显性和流行的CD 8和CD 4表位的疫苗。

DESCRIPTION (provided by applicant): Herpes simplex virus type 1 (HSV-1) is a medically significant pathogen. 60% of US citizens are infected and rates are higher in developing countries. Serious complications, such as permanent brain damage from encephalitis in adults or neonates, and corneal inflammation leading to blindness and poor long- term outcomes even after corneal transplantation, occur at a low per-person rate. Because infection is so prevalent, the cumulative medical impact of these complications is quite considerable. There is no licensed vaccine for HSV-1. A vaccine for HSV-2, a pathogen with overall 50% amino acid identity, had no clinical activity in a recent phase III trial. This HSV-2 vaccine targeted antibody and CD4 T-cell responses, but not CD8 T-cell responses. The long-term goal of this research is the rational design of preventative and therapeutic vaccines for HSV-1. CD8 T-cells are a critical component of the immune response to HSV-1 and localize to infected skin, cornea, retina, and trigeminal ganglia. By implication from human HSV-2 and murine HSV-1 data, CD8 cells persist at sites of healed skin infection to provide early surveillance for reactivation. Almost nothing is known about their fine specificity in humans. We have developed novel technology what allows us to study the CD8 and CD4 T-cell response to HSV-1 in unprecedented detail, including the study of peripheral blood from person who never have herpetic lesions, that is, person with immunologic success. Aim 1 uses a sequence of novel methods: efficient cross-presentation, CD137-based sorting, and a library of cloned HSV-1 ORFs, to decode the CD8 response to HSV with high throughput and definition. We will compare immune responses in persons with symptomatic and asymptomatic HSV-1 infection and identify antigens that have both high population prevalence and correlate with asymptomatic infection. HSV-1-specific CD4s localize to infected eyes, TG, and skin in humans, and to infected tissues in mice, and likely provide important direct antiviral and helper functions. Aim 2 defines with high throughput, using the complete ORFeome, the breadth, specificity, and within-subject true immunodominance hierarchy of HSV-1-specific CD4 responses. The workflow uses CD137-based sorting to and a highly antigenic HSV-1 ORFeome protein set. Dominance assays are combinatorial in two dimensions (responder cell oligoclonality and matrix ORF pooling) and deliver the fine specificity of ~100 individual HSV-1-specific CD4 T-cells per subject to define the CD4 dominance architecture. The overall goal of the Project is to identify HSV-1 ORFs for vaccines that stimulate dominant and prevalent CD8 and CD4 epitopes to persons of diverse HLA type that are correlated with mild infection.