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

描述（由申请人提供）：1型单纯疱疹病毒（HSV-1）是医学上重要的病原体。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引起的永久性脑损伤，以及角膜炎症导致失明和即使在角膜移植后长期预后不良，人均发生率很低。由于感染如此普遍，这些并发症的累积医疗影响相当可观。目前还没有针对1型单纯疱疹病毒的许可疫苗。在最近的一项三期试验中，一种针对HSV-2的疫苗没有临床活性。HSV-2是一种总体上具有50%氨基酸同源性的病原体。这种HSV-2疫苗针对的是抗体和CD4 t细胞应答，而不是CD8 t细胞应答。本研究的长期目标是合理设计1型单纯疱疹病毒的预防和治疗疫苗。CD8 t细胞是HSV-1免疫反应的重要组成部分，可定位于受感染的皮肤、角膜、视网膜和三叉神经节。根据人类HSV-2和小鼠HSV-1的数据，CD8细胞在愈合的皮肤感染部位持续存在，为再激活提供了早期监测。人们对它们在人类身上的精细特异性几乎一无所知。我们已经开发了一种新技术，使我们能够以前所未有的细节研究CD8和CD4 t细胞对HSV-1的反应，包括研究从未患过疱疹病变的人的外周血，也就是免疫成功的人。Aim 1使用一系列新方法：高效交叉呈现、基于cd137的分类和克隆HSV-1 orf库，以高通量和高清晰度解码CD8对HSV的反应。我们将比较有症状和无症状的1型单纯疱疹病毒感染者的免疫反应，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CD4s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。Aim 2使用完整的ORFeome，以高通量定义hsv -1特异性CD4反应的广度、特异性和受试者内真实免疫优势等级。该工作流程使用基于cd137的分选和高抗原HSV-1 ORFeome蛋白集。显性分析在两个维度（应答细胞寡克隆性和基质ORF池化）上进行组合，并提供每个受试者约100个hsv -1特异性CD4 t细胞的良好特异性，以确定CD4显性结构。该项目的总体目标是确定HSV-1 orf疫苗，刺激与轻度感染相关的不同HLA类型的人的显性和流行CD8和CD4表位。

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.